Scientific Bibliography
Reviews
2005 2004 2003 2002 2001 2000
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2005
Mapping molecular networks using proteomics: a vision for patient-tailored combination therapy.
Petricoin EF 3rd, Bichsel VE, Calvert VS, Espina V, Winters M, Young L, Belluco C, Trock BJ, Lippman M, Fishman DA, Sgroi DC, Munson PJ, Esserman LJ, Liotta LA.
J Clin Oncol. 2005 May 20;23(15):3614-21.
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Mapping tumor cell protein networks in vivo will be critical for realizing the promise of patient-tailored molecular therapy. Cancer can be defined as a dysregulation or hyperactivity in the network of intracellular and extracellular signaling cascades. These protein signaling circuits are the ultimate targets of molecular therapy. Each patient's tumor may be driven by a distinct series of molecular pathogenic defects. Thus, for any single molecular targeted therapy, only a subset of cancer patients may respond. Individualization of therapy, which tailors a therapeutic regimen to a tumor molecular portrait, may be the solution to this dilemma. Until recently, the field lacked the technology for molecular profiling at the genomic and proteomic level. Emerging proteomic technology, used concomitantly with genomic analysis, promises to meet this need and bring to reality the clinical adoption of molecular stratification. The activation state of kinase-driven signal networks contains important information relative to cancer pathogenesis and therapeutic target selection. Proteomic technology offers a means to quantify the state of kinase pathways, and provides post-translational phosphorylation data not obtainable by gene arrays. Case studies using clinical research specimens are provided to show the feasibility of generating the critical information needed to individualize therapy. Such technology can reveal potential new pathway interconnections, including differences between primary and metastatic lesions. We provide a vision for individualized combinatorial therapy based on proteomic mapping of phosphorylation end points in clinical tissue material.
Back to Top The molecular make-up of a tumour: proteomics in cancer research.
Kolch W, Mischak H, Pitt AR.
Clin Sci (Lond). 2005 May;108(5):369-83.
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The enormous progress in proteomics, enabled by recent advances in MS (mass spectrometry), has brought protein analysis back into the limelight of cancer research, reviving old areas as well as opening new fields of study. In this review, we discuss the basic features of proteomic technologies, including the basics of MS, and we consider the main current applications and challenges of proteomics in cancer research, including (i) protein expression profiling of tumours, tumour fluids and tumour cells; (ii) protein microarrays; (iii) mapping of cancer signalling pathways; (iv) pharmacoproteomics; (v) biomarkers for diagnosis, staging and monitoring of the disease and therapeutic response; and (vi) the immune response to cancer. All these applications continue to benefit from further technological advances, such as the development of quantitative proteomics methods, high-resolution, high-speed and high-sensitivity MS, functional protein assays, and advanced bioinformatics for data handling and interpretation. A major challenge will be the integration of proteomics with genomics and metabolomics data and their functional interpretation in conjunction with clinical results and epidemiology.
How molecular profiling could revolutionize drug discovery.
Stoughton RB, Friend SH.
Nat Rev Drug Discov. 2005 Apr;4(4):345-50.
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Information from genomic, proteomic and metabolomic measurements has already benefited target discovery and validation, assessment of efficacy and toxicity of compounds, identification of disease subgroups and the prediction of responses of individual patients. Greater benefits can be expected from the application of these technologies on a significantly larger scale; by simultaneously collecting diverse measurements from the same subjects or cell cultures; by exploiting the steadily improving quantitative accuracy of the technologies; and by interpreting the emerging data in the context of underlying biological models of increasing sophistication. The benefits of applying molecular profiling to drug discovery and development will include much lower failure rates at all stages of the drug development pipeline, faster progression from discovery through to clinical trials and more successful therapies for patient subgroups. Upheavals in existing organizational structures in the current 'conveyor belt' models of drug discovery might be required to take full advantage of these methods.
Biomarkers: mining the biofluid proteome.
Veenstra TD, Conrads TP, Hood BL, Avellino AM, Ellenbogen RG, Morrison RS.
Mol Cell Proteomics. 2005 Apr;4(4):409-18.
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Proteomics has brought with it the hope of identifying novel biomarkers for diseases such as cancer. This hope is built on the ability of proteomic technologies, such as mass spectrometry (MS), to identify hundreds of proteins in complex biofluids such as plasma and serum. There are many factors that make this research very challenging beginning with the lack of standardization of sample collection and continuing through the entire analytical process. Fortunately the advances made in the characterization of biofluids using proteomic techniques have been rapid and suggest that these mainly discovery driven approaches will lead to the development of highly specific platforms for diagnosing diseases and monitoring responses to different treatments in the near future.
Informatics for protein identification by mass spectrometry.
Johnson RS, Davis MT, Taylor JA, Patterson SD.
Methods. 2005 Mar;35(3):223-36.
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High throughput protein analysis (i.e., proteomics) first became possible when sensitive peptide mass mapping techniques were developed, thereby allowing for the possibility of identifying and cataloging most 2D gel electrophoresis spots. Shortly thereafter a few groups pioneered the idea of identifying proteins by using peptide tandem mass spectra to search protein sequence databases. Hence, it became possible to identify proteins from very complex mixtures. One drawback to these latter techniques is that it is not entirely straightforward to make matches using tandem mass spectra of peptides that are modified or have sequences that differ slightly from what is present in the sequence database that is being searched. This has been part of the motivation behind automated de novo sequencing programs that attempt to derive a peptide sequence regardless of its presence in a sequence database. The sequence candidates thus generated are then subjected to homology-based database search programs (e.g., BLAST or FASTA). These homology search programs, however, were not developed with mass spectrometry in mind, and it became necessary to make minor modifications such that mass spectrometric ambiguities can be taken into account when comparing query and database sequences. Finally, this review will discuss the important issue of validating protein identifications. All of the search programs will produce a top ranked answer; however, only the credulous are willing to accept them carte blanche.
Proteomics strategies for protein identification.
Resing KA, Ahn NG.
FEBS Lett. 2005 Feb 7;579(4):885-9.
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The information from genome sequencing provides new approaches for systems-wide understanding of protein networks and cellular function. DNA microarray technologies have advanced to the point where nearly complete monitoring of gene expression is feasible in several organisms. An equally important goal is to comprehensive survey cellular proteomes and profile protein changes under different cellular states. This presents a complex analytical problem, due to the chemical variability between proteins and peptides. Here, we discuss strategies to improve accuracy and sensitivity of peptide identification, distinguish represented protein isoforms, and quantify relative changes in protein abundance.
2004
Genomic and proteomic technologies for individualisation and improvement of cancer treatment.
Wulfkuhle J, Espina V, Liotta L, Petricoin E.
Eur J Cancer. 2004 Nov;40(17):2623-32.
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The development of microarray-based technologies for characterising tumours, both at the genomic and proteomic levels, has had a significant impact on the field of oncology. Gene expression profiling of various human tumour tissues has led to the identification of expression patterns related to disease outcome and drug resistance, as well as to the discovery of new therapeutic targets and insights into disease pathogenesis. Protein microarray technologies, such as reverse-phase protein arrays, provide the unique opportunity to profile tissues and assess the activity of signalling pathways within isolated cell populations. This technology can be used to identify patients likely to benefit from specific treatment modalities and also to monitor therapeutic response in samples obtained during and after treatment. Routine application of genomic and proteomic microarray technologies in clinical practice will require significant efforts to standardise the techniques, controls and reference standards, and analytical tools used. Extensive, independent validation using large, statistically-powered datasets will also be necessary. Inclusion of concomitant genomic and proteomic-based molecular profiling techniques into clinical trial protocols will bring us closer to the reality of patient-tailored therapy.
Strategies for shotgun identification of post-translational modifications by mass spectrometry.
Cantin GT, Yates JR 3rd.
J Chromatogr A. 2004 Oct 22;1053(1-2):7-14.
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The global identification of post-translationally modified proteins is a difficult challenge that is currently being addressed by many researchers in the field of mass spectrometry (MS)-based proteomics. The ability to identify thousands of proteins by shotgun-based strategies has made the mere idea of a global analysis of a particular protein modification seem reasonable. There has been much progress in the development of methods that make use of shotgun-based protein identification in the analysis of a wide variety of protein modifications, some of which will be discussed here.
Proteomic patterns for early cancer detection.
Veenstra TD, Prieto DA, Conrads TP.
Drug Discov Today. 2004 Oct 15;9(20):889-97.
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The advent of proteomics has brought with it the hope of discovering novel biomarkers that can be used to diagnose diseases, predict susceptibility, and monitor progression. Much of this effort has focused on the mass spectral identification of the thousands of proteins that populate complex biosystems such as serum and tissues. A revolutionary approach in proteomic pattern analysis has emerged as an effective method for the early diagnosis of diseases such as ovarian, breast, and prostate cancer. This technology is capable of analyzing hundreds of clinical samples per day and has the potential to be a
novel, highly sensitive diagnostic tool for the early detection of diseases, or as a predictor of response to therapy.
Turning protein crystallisation from an art into a science.
Chayen NE.
Curr Opin Struct Biol. 2004 Oct;14(5):577-83.
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Protein crystallisation has gained a new strategic and commercial relevance in the post-genomic era because of its pivotal role in structural genomics. Producing high-quality crystals has always been a bottleneck to structure determination and, with the advent of proteomics, this problem is becoming increasingly acute. The task of producing suitable crystals may be tackled using two approaches. The first relies on empirical techniques that are based mainly on trial and error, and what is perceived to be the 'art' of crystallisation. The second approach is aimed at gaining an understanding of the fundamental principles that govern crystallisation; this knowledge may be applied to design experimental methodology for producing high-quality crystals of medical and industrial interest.
A vision for the National Cancer Program in the United States.
von Eschenbach AC.
Nat Rev Cancer. 2004 Oct;4(10):820-8.
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The intersection of two noble endeavors - the scientists' quest to understand life itself and the physicians' dedication to relieve suffering and prolong life - came into sharp focus in 1971 with the United States National Cancer Act. This focus has led to an exponential expansion of our understanding of cancer at the genetic, molecular and cellular levels, and concomitant advances in our ability to disrupt the disease process through prevention, early detection and successful treatment. At the National Cancer Institute we are committed to capitalize on these achievements. A new era is now within our grasp, a time when no one suffers or dies as a result of cancer.
Miniaturized proteomics and peptidomics using capillary liquid separation and high resolution mass spectrometry.
Ramstrom M, Bergquist J.
FEBS Lett. 2004 Jun 1;567(1):92-5.
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Knowledge of the protein and peptide content in a tissue or a body fluid is vital in many areas of medical and biomedical sciences. Information from proteomic and peptidomic studies may reveal alterations in expression due to, e.g., a disease and facilitate the understanding of the pathophysiology and the identification of biological markers. In this minireview, we discuss miniaturized proteomic and peptidomic approaches that have been applied in our laboratory in order to investigate the protein and peptide contents of body fluids (such as plasma, cerebrospinal and amniotic fluid), as well as extracted tissues. The methods involve miniaturized liquid separation, i.e., capillary liquid chromatography and capillary electrophoresis, combined with high resolution mass spectrometry (MS), i.e., Fourier transform ion cyclotron resonance MS. These approaches provide the opportunity to analyze samples of small volumes with high throughput, high sensitivity, good dynamic range and minimal sample handling. Also, the experiments are relatively easy to automate.
Prediction of post-translational glycosylation and phosphorylation of proteins from the amino acid sequence.
Blom N, Sicheritz-Ponten T, Gupta R, Gammeltoft S, Brunak S.
Proteomics. 2004 Jun;4(6):1633-49.
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Post-translational modifications (PTMs) occur on almost all proteins analyzed to date. The function of a modified protein is often strongly affected by these modifications and therefore increased knowledge about the potential PTMs of a target protein may increase our understanding of the molecular processes in which it takes part. High-throughput methods for the identification of PTMs are being developed, in particular within the fields of proteomics and mass spectrometry. However, these methods are still in their early stages, and it is indeed advantageous to cut down on the number of experimental steps by integrating computational approaches into the validation procedures. Many advanced methods for the prediction of PTMs exist and many are made publicly available. We describe our experiences with the development of prediction methods for phosphorylation and glycosylation sites and the development of PTM-specific databases. In addition, we discuss novel ideas for PTM visualization (exemplified by kinase landscapes) and improvements for prediction specificity (by using ESS--evolutionary stable sites). As an example, we present a new method for kinase-specific prediction of phosphorylation sites, NetPhosK, which extends our earlier and more general tool, NetPhos. The new server, NetPhosK, is made publicly available at the URL http://www.cbs.dtu.dk/services/NetPhosK/. The issues of underestimation, over-prediction and strategies for improving prediction specificity are also discussed.
Quantification in proteomics through stable isotope coding: a review.
Julka S, Regnier F.
J Proteome Res. 2004 May-Jun;3(3):350-63.
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This review focuses on techniques for quantification and identification in proteomics by stable isotope coding. Methods are examined for analyzing expression, post-translational modifications, protein:protein interactions, single amino acid polymorphism, and absolute quantification. The bulk of the quantification literature in proteomics focuses on expression analysis, where a wide variety of methods targeting different features of proteins are described. Methods for the analysis of post-translational modification (PTM) focus primarily on phosphorylation and glycosylation, where quantification is achieved in two ways, either by substitution or tagging of the PTM with an isotopically coded derivatizing agent in a single process or by coding and selecting PTM modified peptides in separate operations. Absolute quantification has been achieved by age-old internal standard methods, in which an isotopically labeled isoform of an analyte is synthesized and added to a mixture at a known concentration. One of the surprises is that isotope coding can be a valuable aid in the examination of intermolecular association of proteins through stimulus:response studies. Preliminary efforts to recognize single amino acid polymorphism are also described. The review ends with the conclusion that (1) isotope ratio analysis of protein concentration between samples does not necessarily relate directly to protein expression and rate of PTM and (2) that multiple new methods must be developed and applied simultaneously to make existing stable isotope quantification methods more meaningful. Although stable isotope coding is a powerful, wonderful new technique, multiple analytical issues must be solved for the technique to reach its full potential as a tool to study biological systems.
Metabonomics: systems biology in pharmaceutical research and development.
Lindon JC, Holmes E, Nicholson JK.
Curr Opin Mol Ther. 2004 Jun;6(3):265-72.
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Metabonomics uses a combination of data-rich analytical chemical methods such as nuclear magnetic resonance spectroscopy and mass spectrometry together with chemometrics for profiling metabolism and interpreting metabolic fingerprints in complex biological systems. The methods have been applied in many areas of relevance to pharmaceutical research and development, including drug safety assessment, characterization of genetically modified animal models of disease, diagnosis of human disease, understanding physiological variation and drug therapy monitoring. As well as providing a novel means of sample classification and effect evaluation, the approach can lead to identification of combinations of biomarkers for those effects. These attributes mean that metabonomics will be integral in the drive towards personalized healthcare.
MALDI: more than peptide mass fingerprints.
Stuhler K, Meyer HE.
Curr Opin Mol Ther. 2004 Jun;6(3):239-48.
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Over the past decade, matrix-assisted laser desorption/ionization (MALDI) has developed from a phenomenon that was interesting only to physicists and mass spectrometrists, and has evolved into a widespread applied ionization technique in chemistry, biology and biomedicine. The introduction of MALDI offers all the advantages of mass spectrometry (MS) for the analysis of large biomolecules. Proteins, carbohydrates and oligonucleotides have been analyzed with MS-inherent accuracy, sensitivity, resolution and speed. In addition to electrospray ionization-MS, MALDI-MS has a great impact on the challenges of the post-genome area. The recent status of MALDI application in proteomics, biomolecular interaction analysis-MS and carbohydrate analysis, in addition to single nucleotide polymorphism genotyping is reviewed.
Combinatorial approaches to protein stability and structure.
Magliery TJ, Regan L.
Eur J Biochem. 2004 May;271(9):1595-608.
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Why do proteins adopt the conformations that they do, and what determines their stabilities? While we have come to some understanding of the forces that underlie protein architecture, a precise, predictive, physicochemical explanation is still elusive. Two obstacles to addressing these questions are the unfathomable vastness of protein sequence space, and the difficulty in making direct physical measurements on large numbers of protein variants. Here, we review combinatorial methods that have been applied to problems in protein biophysics over the last 15 years. The effects of hydrophobic core composition, the most important determinant of structure and stability, are still poorly understood. Particular attention is given to core composition as addressed by library methods. Increasingly useful screens and selections, in combination with modern high-throughput approaches borrowed from genomics and proteomics efforts, are making the empirical, statistical correlation between sequence and structure a tractable problem for the coming years.
HUPO initiatives relevant to clinical proteomics.
Hanash S.
Mol Cell Proteomics. 2004 Apr;3(4):298-301.
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The past few years have seen a tremendous interest in the potential of proteomics to address unmet needs in biomedicine. Such unmet needs include more effective strategies for early disease detection and monitoring and more effective therapies, in addition to developing a better understanding of disease pathogenesis. Proteomics is particularly suited for investigating biological fluids to identify disease-related alterations and to develop molecular signatures for disease processes. However, much of the effort undertaken in clinical proteomics to date represents either demonstrations of principles or relatively small-scale studies when compared with genomics effort and accomplishments or more pertinently when contrasted with the tremendous untapped potential of clinical proteomics. Clearly, we are in the early stages. What seems to be urgently needed is an organized effort to build a solid foundation for proteomics that includes developing a much needed infrastructure with adequate resources. The Human Proteome Organization (HUPO) is fostering an organized international effort in proteomics that includes initiatives around organ systems and biological fluids that have disease relevance as well as development of proteomics resources.
How industry is approaching the search for new diagnostic markers and biomarkers.
Zolg JW, Langen H.
Mol Cell Proteomics. 2004 Apr;3(4):345-54.
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In the diagnostic and the pharmaceutical industry there is a constant need for new diagnostic markers and biomarkers with improved sensitivity and specificity. During the last 5 years, only a few novel diagnostic markers have been introduced into the market. Proteomics technologies are now offering unique chances to identify new candidate markers. Before a marker can be introduced into the market, three successive developmental phases have to be completed: the discovery phase, in which a variety of proteomics technologies are applied to identify marker candidates; the prototype developmental phase, in which immunological assays are established and validated in defined sample collectives; and finally the product development phase, with assay formats suitable for automated platforms. The hurdles that a potential candidate marker has to pass in each developmental phase before reaching the market are considerable. The costs are increasing from phase to phase, and in industry a number of questions concerning the medical need and the potential return on investment have to be answered before a proteomics discovery project is started. In this review, we will cover aspects of all three developmental phases including the repertoire of discovery tools for protein separation as well as giving an outline of modern principles of mass spectrometry for the identification of proteins.
The role of emerging genomics and proteomics technologies in cancer drug target discovery.
Onyango P.
Curr Cancer Drug Targets. 2004 Mar;4(2):111-24.
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Cancer drugs have traditionally been identified in screens designed to produce broad biological end points such as cell death. A serious undesired outcome of drugs discovered in these screens is that the mechanism of drug action is unknown and such drugs often have adverse side effects. Designing cancer drugs that act on specific targets offer the advantage that the mechanism of drug action can be understood and accurately monitored in clinical trials leading to development of better drugs. The pharmacological industry has recently shifted to a target directed drug discovery model. However, until recently potential cancer drug targets comprised of only a small fraction of the human genome. The human genome project and high-throughput structural and functional genomics have dramatically increased the number of cancer drug targets. Deciphering cancer drug targets requires the understanding of biochemical pathways that are affected in the cancer genome. It has been suggested that utilization of Single-nucleotide polymorphisms (SNPs) will aid in identifying individuals at high risk of developing certain cancers, and will also help in development of tailored medication or identify genetic profiles of specific drug action and toxicity. Achieving successful new cancer drug development schemes will require a merger of research disciplines that include pharmacology, genomics, comparative genomics, functional genomics, proteomics and bioinformatics. In this review the significance and challenges of these rapidly evolving technologies in cancer drug target discovery are discussed.
Protein nanocrystallography: a new approach to structural proteomics.
Pechkova E, Nicolini C.
Trends Biotechnol. 2004 Mar;22(3):117-22.
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This article describes a new approach to structural proteomics that can produce and characterize diffracting, stable and radiation-resistant crystals of miniscule dimensions using nanotechnology. We believe that the protein microcrystals obtained by nanotechnology-based protein thin-film template crystallization, as well as groundbreaking technology, such as atomic force microscopy, nanogravimetry and synchrotron microfocus, have enabled protein nanocrystallography to be defined as a unique technology capable of forming and characterizing stable protein microcrystals down to atomic resolution. A new route from art to science and technology has, therefore, been opened in protein crystallography, and it could be used to unravel the mysteries of many systems that remain unsolved.
Bioinformatics in proteomics.
Blueggel M, Chamrad D, Meyer HE.
Curr Pharm Biotechnol. 2004 Feb;5(1):79-88.
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Proteomics technologies are under continuous improvements and new technologies are introduced. Nowadays high throughput acquisition of proteome data is possible. The young and rapidly emerging field of bioinformatics in proteomics is introducing new algorithms to handle large and heterogeneous data sets and to improve the knowledge discovery process. For example new algorithms for image analysis of two dimensional gels have been developed within the last five years. Within mass spectrometry data analysis algorithms for peptide mass fingerprinting (PMF) and peptide fragmentation fingerprinting (PFF) have been developed. Local proteomics bioinformatics platforms emerge as data management systems and knowledge bases in Proteomics. We review recent developments in bioinformatics for proteomics with emphasis on expression proteomics.
Molecular therapeutics: promise and challenges.
Kohn EC, Lu Y, Wang H, Yu Q, Yu S, Hall H, Smith DL, Meric-Bernstam F, Hortobagyi GN, Mills GB.
Semin Oncol. 2004 Feb;31(1 Suppl 3):39-53.
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The ability to analyze the genetic and epigenetic aberrations present in a particular patient's tumor on a global basis is rapidly maturing. The emerging fields of functional genomics and functional proteomics offer the opportunity to translate these advances into a full comprehension of the pathophysiology of cancer. Linking these approaches to chemical genomics and molecular therapeutics should provide an expanding repertoire of targeted therapeutics for clinical evaluation. Novel clinical trial designs that can determine the efficacy of targeted therapeutics in patients selected for aberrations in the target are needed to evaluate the wealth of new drugs becoming available. The promise of these technologies and advances in our understanding of cancer is immense, making it our responsibility to see them to fruition. These technologies should lead to a new era of individualized molecular medicine, wherein we will treat each patient with a "prescription" based on the genetic changes in each patient's tumor and their own genetic make-up, resulting in more effective and less toxic therapy.
Enhanced functional information from predicted protein networks.
McDermott J, Samudrala R.
Trends Biotechnol. 2004 Feb;22(2):60-2; discussion 62-3.
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Experimentally derived genome-wide protein interaction networks have been useful in the elucidation of functional information that is not evident from examining individual proteins but determination of these networks is complex and time consuming. To address this problem, several computational methods for predicting protein networks in novel genomes have been developed. A recent publication by Date and Marcotte describes the use of phylogenetic profiling for elucidating novel pathways in proteomes that have not been experimentally characterized. This method, in combination with other computational methods for generating protein-interaction networks, might help identify novel functional pathways and enhance functional annotation of individual proteins.
Tagged library approach to chemical genomics and proteomics.
Mitsopoulos G, Walsh DP, Chang YT.
Curr Opin Chem Biol. 2004 Feb;8(1):26-32.
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Proteomics and chemical genomics face great challenges in the form of molecular libraries of ever increasing size and diversity requiring rapid screening, coupled with a growing number of target proteins for which complimentary molecular ligands are sought. Proteomics and chemical genomics are at a stage that requires techniques which can dramatically accelerate the discovery process. One technique that has shown great promise in accomplishing this is the tagged library approach. It entails the synthetic inclusion of an internal tag from the beginning of the synthesis. This tag adds another degree of functionality to the molecule, in addition to mere ligation, that eliminates the need for time-consuming steps downstream in the process. The tag's functional possibilities span a variety of uses including internal fluorophores, intrinsic binding motifs that enable compound identification, functionalities that play the major role in the synthesis of the ligand itself, and internal linkers that eliminate the need for lengthy 'tether effect' structure-activity relationship studies.
Tracking cell signaling protein expression and phosphorylation by innovative proteomic solutions.
Pelech S.
Curr Pharm Biotechnol. 2004 Feb;5(1):69-77.
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The most challenging and fruitful biomedical research endeavor of this decade will be the mapping of cell signaling systems and establishing their linkages to normal and disease-related processes. Amongst other things, the Human Genome Sequencing Project has greatly facilitated MALDI-TOF mass spectrometry identification of proteins that have been resolved by standard 2D gel electrophoresis. However, the low abundance of protein kinases and other signal transduction proteins has rendered their analyses particularly problematic without some means of purification and enrichment from cell and tissue lysates. Antibodies have been the most specific affinity probes for tracking target proteins, but their variable quality and high cost preclude their deployment in most discovery-based proteomics studies. Current multi-immunoblotting techniques can permit the probing of a single mini-SDS-PAGE gel with 50 or more antibodies at a time to monitor large changes in the expression and phosphorylation states of signaling proteins. The development of new affinity probes to replace antibodies is necessary to drive large scale proteomics studies. Such affinity probes could include short peptide antibody mimetics (PAM's) and oligonucleotide aptamers that when spotted in 2D array formats (e.g. membrane macroarrays, glass microarrays) or presented on specific beads (e.g. Luminex beads) can capture target proteins for their specific enrichment. The bound target proteins can then be detected using reporter antibodies or other specific probes for their quantitation by high throughput systems. These new proteomics methodologies will accelerate assessment of specific protein expression, post-translational modification, protein-protein interactions and protein-drug interactions to provide a more holistic view of cellular operations and how they might be manipulated under pathological circumstances.
Functional protein microarrays: ripe for discovery.
Predki PF.
Curr Opin Chem Biol. 2004 Feb;8(1):8-13.
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The manufacture and use of protein microarrays with correctly folded and functional content presents significant challenges. Despite this, the feasibility and utility of such undertakings are now clear, and exciting progress has recently been demonstrated in the areas of content generation, printing strategies and protein immobilization. More importantly, we are now beginning to enjoy the fruits of these efforts as functional protein microarrays are being increasingly employed for biological discovery purposes. Recent examples of this include the characterization of autoantibody responses, antibody specificity profiling, protein-protein domain interaction profiling and a comprehensive characterization of coiled-coil interactions. The best, however, is yet to come.
Protein-protein interaction networks: from interactions to networks.
Cho S, Park SG, Lee do H, Park BC.
J Biochem Mol Biol. 2004 Jan 31;37(1):45-52.
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The goal of interaction proteomics that studies the protein-protein interactions of all expressed proteins is to understand biological processes that are strictly regulated by these interactions. The availability of entire genome sequences of many organisms and high-throughput analysis tools has led scientists to study the entire proteome (Pandey and Mann, 2000). There are various high-throughput methods for detecting protein interactions such as yeast two-hybrid approach and mass spectrometry to produce vast amounts of data that can be utilized to decipher protein functions in complicated biological networks. In this review, we discuss recent developments in analytical methods for large-scale protein interactions and the future direction of interaction proteomics.
Phylogenomic inference of protein molecular function: advances and challenges.
Sjolander K.
Bioinformatics. 2004 Jan 22;20(2):170-9.
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MOTIVATION: Protein families evolve a multiplicity of functions through gene duplication, speciation and other processes. As a number of studies have shown, standard methods of protein function prediction produce systematic errors on these data. Phylogenomic analysis--combining phylogenetic tree construction, integration of experimental data and differentiation of orthologs and paralogs--has been proposed to address these errors and improve the accuracy of functional classification. The explicit integration of structure prediction and analysis in this framework, which we call structural phylogenomics, provides additional insights into protein superfamily evolution. RESULTS: Results of protein functional classification using phylogenomic analysis show fewer expected false positives overall than when pairwise methods of functional classification are employed. We present an overview of the motivations and fundamental principles of phylogenomic analysis, new methods developed for the key tasks, benchmark datasets for these tasks (when available) and suggest procedures to increase accuracy. We also discuss some of the methods used in the Celera Genomics high-throughput phylogenomic classification of the human genome. AVAILABILITY: Software tools from the Berkeley Phylogenomics Group are available at http://phylogenomics.berkeley.edu.
Genomic and proteomic approaches for studying human cancer: prospects for true patient-tailored therapy.
Carr KM, Rosenblatt K, Petricoin EF, Liotta LA.
Hum Genomics. 2004 Jan;1(2):134-40.
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Global gene expression analysis is beginning to move from the laboratories of basic investigators to large-scale clinical trials. The potential of this technology to improve diagnosis and tailored treatment of human disease may soon be realised, now that several comprehensive studies have demonstrated the utility of gene expression profiles for the classification of tumours into distinct, clinically relevant subtypes and the prediction of clinical outcomes. In addition, new data from the emerging proteomics platforms add another layer of molecular information to the study of human disease, as scientists attempt to catalogue a complete inventory of the proteins encoded by the genome and to establish a 'biosignature' profile of human health and disease. As a result, it is anticipated that, together, these technologies will facilitate the comprehensive study of genes, gene products and signalling pathways so that the objective of personalized molecular medicine can be achieved. This paper will review the studies that best demonstrate how genomics and proteomics technologies can be used to improve cancer diagnosis and treatment it will specifically highlight the important work being incorporated into clinical trials.
Proteomic patterns as a diagnostic tool for early-stage cancer: a review of its progress to a clinically relevant tool.
Conrads TP, Hood BL, Issaq HJ, Veenstra TD.
Mol Diagn. 2004;8(2):77-85.
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The pace of development in novel technologies that promise improvements in the early diagnosis of disease is truly impressive. One such technology at the forefront of this revolution is mass spectrometry. New capabilities in mass spectrometry have provided the means for the development of proteomics, and the race is on to find innovative ways to apply this powerful technology to solving the problems faced in clinical medicine. One area that has garnered much attention over the past few years is the use of mass spectral patterns for cancer diagnostics. The use of these so-called 'proteomic patterns' for disease diagnosis relies fundamentally on the pattern of signals observed within a mass spectrum rather than the more conventional identification and quantitation of a biomarker such as in the case of cancer antigen-125- or prostate-specific antigen. The inherent throughput of proteomic pattern technology enables the analysis of hundreds of clinical samples per day. Currently, there are two primary means by which proteomic patterns can be acquired, surface-enhanced laser desorption/ionization (SELDI) and an electrospray ionization (ESI) method that has been popularized under the name, OvaCheck. In this review, an historical perspective on the development of proteomic patterns for the diagnosis of early-stage cancers is described. In addition, a critical assessment of the overall technology is presented with an emphasis on the steps required to enable proteomic pattern analysis to become a viable clinical tool for diagnosing early-stage cancers.
Structural characterization of proteins and peptides.
Deutzmann R.
Methods Mol Med. 2004;94:269-97.
[ expand abstract ]
The primary structure of proteins is nowadays determined by DNA sequencing, and a variety of genomes are already known. Nevertheless, protein sequencing/identification is still indispensable to analyze the proteins expressed in a cell, to identify specific proteins, and to determine posttranslational modifications. Proteins of interest are typically available in low microgram amounts or even less. The separation method of choice is gel electrophoresis, followed by blotting to PVDF membrane for N-terminal sequencing or by in-gel digestion to generate peptides that can be separated by HPLC. Structural analysis can be done by Edman degradation or mass spectrometry (MS). Edman degradation is the older method based on successive removal of N-terminal amino acids by chemical methods. Sequencing of a peptide requires many hours, the sensitivity is in the range of 2-5 pmol of a purified peptide. Nevertheless, Edman degradation is still the workhorse in the lab for routine work such as identification of blotted proteins. It is also the method of choice for sequencing unknown proteins/ peptides and modified peptides. MS has routinely been used with peptides in the range of 100 fmol or even less. In contrast to Edman degradation, complex mixtures such as tryptic digests can be analyzed, making HPLC separation of peptides unnecessary. MS is a very fast method that can be automated. It is the method of choice for sensitive analysis and large-scale applications (proteomics). Two different ionization methods are commonly used to generate peptide/protein ions for MS analysis. These are MALDI (matrix assisted laser desorption and ionization) and ESI (electrospray ionization). They can be combined with a variety of mass analyzers (TOF, quadrupole, ion trap). Proteins are either identified by searching databases with the masses of proteolytic peptides (peptide mass fingerprinting) or using fragmentation data (raw MS/MS spectra or sequence tags). This approach requires that the protein is known and listed in the database. De novo sequencing by MS of peptides is possible, but very time consuming and not a routine application, in contrast to Edman degradation. The aim of this chapter is to introduce to basic theory, practical applications and limitations of the various methods, to enable the non-expert scientist to decide which method is best suited for his project and which kind of sample preparation is necessary.
Use of a small molecule-based affinity system for the preparation of protein microarrays.
Hughes KA.
Methods Mol Biol. 2004;264:111-21.
[ expand abstract ]
This chapter describes a method for preparing protein microarrays, using a small-molecule, chemical affinity system.
2003
Microfluidic systems in proteomics.
Lion N, Rohner TC, Dayon L, Arnaud IL, Damoc E, Youhnovski N, Wu ZY, Roussel C, Josserand J, Jensen H, Rossier JS, Przybylski M, Girault HH.
Electrophoresis. 2003 Nov;24(21):3533-62.
[ expand abstract ]
We present the state-of-the-art in miniaturized sample preparation, immunoassays, one-dimensional and multidimensional analyte separations, and coupling of microdevices with electrospray ionization-mass spectrometry. Hyphenation of these different techniques and their relevance to proteomics will be discussed. In particular, we will show that analytical performances of microfluidic analytical systems are already close to fulfill the requirements for proteomics, and that miniaturization results at the same time in a dramatic increase in analysis throughput. Throughout this review, some examples of analytical operations that cannot be achieved without microfluidics will be emphasized. Finally, conditions for the spreading of microanalytical systems in routine proteomic labs will be discussed.
Functional genomics and proteomics as a foundation for systems biology.
Aggarwal K, Lee KH.
Brief Funct Genomic Proteomic. 2003 Oct;2(3):175-84.
[ expand abstract ]
Developments in high-throughput measurement technologies for biological molecules have created a paradigm shift in modern life science research. The field of systems biology attempts to provide a systems-level understanding by systematically organising the genomic, functional genomic and proteomic data obtained from genetic and environmental perturbations of interest and using the data to build a descriptive and mechanistic model of the biological phenomena. The goal is to build a mathematical framework with some predictive abilities. This review highlights the need for system-level understanding, lists some of the high-throughput measurement tools of importance in systems biology, reviews various types of experimental and computational approaches being used in systems biology research and attempts to address some of the challenges facing this research community.
Integrating 'omic' information: a bridge between genomics and systems biology.
Ge H, Walhout AJ, Vidal M.
Trends Genet. 2003 Oct;19(10):551-60.
[ expand abstract ]
The availability of genome sequences for several organisms, including humans, and the resulting first-approximation lists of genes, have allowed a transition from molecular biology to 'modular biology'. In modular biology, biological processes of interest, or modules, are studied as complex systems of functionally interacting macromolecules. Functional genomic and proteomic ('omic') approaches can be helpful to accelerate the identification of the genes and gene products involved in particular modules, and to describe the functional relationships between them. However, the data emerging from individual omic approaches should be viewed with caution because of the occurrence of false-negative and false-positive results and because single annotations are not sufficient for an understanding of gene function. To increase the reliability of gene function annotation, multiple independent datasets need to be integrated. Here, we review the recent development of strategies for such integration and we argue that these will be important for a systems approach to modular biology.
Application of proteomics for discovery of protein biomarkers.
Hale JE, Gelfanova V, Ludwig JR, Knierman MD.
Brief Funct Genomic Proteomic. 2003 Oct;2(3):185-93.
[ expand abstract ]
Biomarkers of drug efficacy and toxicity are becoming a key need in the drug development process. Mass spectral-based proteomic technologies are ideally suited for the discovery of protein biomarkers in the absence of any prior knowledge of quantitative changes in protein levels. The success of any biomarker discovery effort will depend upon the quality of samples analysed, the ability to generate quantitative information on relative protein levels and the ability to readily interpret the data generated. This review will focus on the strengths and weaknesses of technologies currently utilised to address these issues.
Shotgun proteomics: integrating technologies to answer biological questions.
McDonald WH, Yates JR 3rd.
Curr Opin Mol Ther. 2003 Jun;5(3):302-9.
[ expand abstract ]
Proteomics is providing us with a variety of exciting new strategies to address biological questions. These strategies must integrate a series of seperative and analytical technologies to deal with the immense complexity involved. At some point in the process, the proteins are usually digested with a proteolytic enzyme to generate shorter peptides that are more easily analyzed by mass spectrometry. Shotgun proteomics relies on separation after this digestion step and takes advantage of tandem mass spectrometry to infer the amino acid sequence of individual peptides. Advances in quantitation, and the ability to find sites of post-translational modification are expanding the scope of questions that can be asked. The ultimate success of any proteomic experiment is dictated not only by an appropriate choice of seperative and analytical techniques, but also by making certain that the biological aspects of the experiment are focused and well designed.
Mass spectrometry-based proteomics.
Aebersold R, Mann M.
Nature. 2003 Mar 13;422(6928):198-207.
[ expand abstract ]
Recent successes illustrate the role of mass spectrometry-based proteomics as an indispensable tool for molecular and cellular biology and for the emerging field of systems biology. These include the study of protein-protein interactions via affinity-based isolations on a small and proteome-wide scale, the mapping of numerous organelles, the concurrent description of the malaria parasite genome and proteome, and the generation of quantitative protein profiles from diverse species. The ability of mass spectrometry to identify and, increasingly, to precisely quantify thousands of proteins from complex samples can be expected to impact broadly on biology and medicine.
Biomedical informatics for proteomics.
Boguski MS, McIntosh MW.
Nature. 2003 Mar 13;422(6928):233-7.
[ expand abstract ]
Success in proteomics depends upon careful study design and high-quality biological samples. Advanced information technologies, and also an ability to use existing knowledge to the full, will be crucial in making sense of the data. Despite its genome-scale potential, proteome analysis is at a much earlier stage of development than genomics and gene expression (microarray) studies. Fundamental issues involving biological variability, pre-analytic factors and analytical reproducibility remain to be resolved. Consequently, the analysis of proteomics data is currently informal and relies heavily on expert opinion. Databases and software tools developed for the analysis of molecular sequences and microarrays are helpful, but are limited owing to the unique attributes of proteomics data and differing research goals.
Proteomic analysis of post-translational modifications.
Mann M, Jensen ON.
Nat Biotechnol. 2003 Mar;21(3):255-61.
[ expand abstract ]
Post-translational modifications modulate the activity of most eukaryote proteins. Analysis of these modifications presents formidable challenges but their determination generates indispensable insight into biological function. Strategies developed to characterize individual proteins are now systematically applied to protein populations. The combination of function- or structure-based purification of modified 'subproteomes', such as phosphorylated proteins or modified membrane proteins, with mass spectrometry is proving particularly successful. To map modification sites in molecular detail, novel mass spectrometric peptide sequencing and analysis technologies hold tremendous potential. Finally, stable isotope labeling strategies in combination with mass spectrometry have been applied successfully to study the dynamics of modifications.
Proteomics: the first decade and beyond.
Patterson SD, Aebersold RH.
Nat Genet. 2003 Mar;33 Suppl:311-23.
[ expand abstract ]
Proteomics is the systematic study of the many and diverse properties of proteins in a parallel manner with the aim of providing detailed descriptions of the structure, function and control of biological systems in health and disease. Advances in methods and technologies have catalyzed an expansion of the scope of biological studies from the reductionist biochemical analysis of single proteins to proteome-wide measurements. Proteomics and other complementary analysis methods are essential components of the emerging 'systems biology' approach that seeks to comprehensively describe biological systems through integration of diverse types of data and, in the future, to ultimately allow computational simulations of complex biological systems.
Proteomics in translational cancer research: toward an integrated approach.
Celis JE, Gromov P.
Cancer Cell. 2003 Jan;3(1):9-15.
[ expand abstract ]
Proteomics provides powerful tools for the study of clinically relevant samples in the context of translational cancer research. Here we briefly review applications of gel-based proteomics for the study of bladder and lung cancer using fresh tissue biopsies. In general, these studies have emphasized the potential of the technology for biomarker discovery, as well as for addressing the issue of cancer heterogeneity.
Proteomic approaches to the diagnosis, treatment, and monitoring of cancer.
Wulfkuhle JD, Paweletz CP, Steeg PS, Petricoin EF 3rd, Liotta L.
Adv Exp Med Biol. 2003;532:59-68.
[ expand abstract ]
The field of proteomics holds promise for the discovery of new biomarkers for the early detection and diagnosis of disease, molecular targets for therapy and markers for therapeutic efficacy and toxicity. A variety of proteomics approaches may be used to address these goals. Two-dimensional gel electrophoresis (2D-PAGE) is the cornerstone of many discovery-based proteomics studies. Technologies such as laser capture microdissection (LCM) and highly sensitive MS methods are currently being used together to identify greater numbers of lower abundance proteins that are differentially expressed between defined cell populations. Newer technologies such as reverse phase protein arrays will enable the identification and profiling of target pathways in small biopsy specimens. Surface-enhanced laser desorption/ionization time-of-flight (SELDI-TOF) analysis enables the high throughput characterization of lysates from very few tumor cells or body fluids and may be best suited for diagnosis and monitoring of disease. Such technologies are expected to supplement our arsenal of mRNA-based assays, and we believe that in the future, entire cellular networks and not just a single deregulated protein will be the target of therapeutics and that we will soon be able to monitor the status of these pathways in diseased cells before, during and after therapy.
2002
Proteomics and models for enzyme cooperativity.
Koshland DE Jr, Hamadani K.
J Biol Chem. 2002 Dec 6;277(49):46841-4.
[ expand abstract ]
(No Abstract)
The structure of the protein universe and genome evolution.
Koonin EV, Wolf YI, Karev GP.
Nature. 2002 Nov 14;420(6912):218-23.
[ expand abstract ]
Despite the practically unlimited number of possible protein sequences, the number of basic shapes in which proteins fold seems not only to be finite, but also to be relatively small, with probably no more than 10,000 folds in existence. Moreover, the distribution of proteins among these folds is highly non-homogeneous -- some folds and superfamilies are extremely abundant, but most are rare. Protein folds and families encoded in diverse genomes show similar size distributions with notable mathematical properties, which also extend to the number of connections between domains in multidomain proteins. All these distributions follow asymptotic power laws, such as have been identified in a wide variety of biological and physical systems, and which are typically associated with scale-free networks. These findings suggest that genome evolution is driven by extremely general mechanisms based on the preferential attachment principle.
The human plasma proteome: history, character, and diagnostic prospects.
Anderson NL, Anderson NG.
Mol Cell Proteomics. 2002 Nov;1(11):845-67.
[ expand abstract ]
The human plasma proteome holds the promise of a revolution in disease diagnosis and therapeutic monitoring provided that major challenges in proteomics and related disciplines can be addressed. Plasma is not only the primary clinical specimen but also represents the largest and deepest version of the human proteome present in any sample: in addition to the classical "plasma proteins," it contains all tissue proteins (as leakage markers) plus very numerous distinct immunoglobulin sequences, and it has an extraordinary dynamic range in that more than 10 orders of magnitude in concentration separate albumin and the rarest proteins now measured clinically. Although the restricted dynamic range of conventional proteomic technology (two-dimensional gels and mass spectrometry) has limited its contribution to the list of 289 proteins (tabulated here) that have been reported in plasma to date, very recent advances in multidimensional survey techniques promise at least double this number in the near future. Abundant scientific evidence, from proteomics and other disciplines, suggests that among these are proteins whose abundances and structures change in ways indicative of many, if not most, human diseases. Nevertheless, only a handful of proteins are currently used in routine clinical diagnosis, and the rate of introduction of new protein tests approved by the United States Food and Drug Administration (FDA) has paradoxically declined over the last decade to less than one new protein diagnostic marker per year. We speculate on the reasons behind this large discrepancy between the expectations arising from proteomics and the realities of clinical diagnostics and suggest approaches by which protein-disease associations may be more effectively translated into diagnostic tools in the future.
Integrated approaches to therapeutic target gene discovery.
DeFife KM, Wong-Staal F.
Curr Opin Drug Discov Devel. 2002 Sep;5(5):683-9 .
[ expand abstract ]
The identification of tractable drug targets is the first critical step in the long process of drug development, and the challenge for scientists in the post-genomic era is to couple gene and protein sequence information with biological insight to identify genes with the greatest therapeutic and commercial potential. An extraordinary array of genomics- and proteomics-based techniques is available for this endeavor, and combining multiple, complementary approaches enhances the informative power of such experimentation.
Functional proteomics: The goalposts are moving.
Hubbard MJ.
Proteomics. 2002 Sep;2(9):1069-78.
[ expand abstract ]
Holistic understanding of protein function is a primary goal of the post-genome sequencing era. Functional genomic approaches are powerful and relatively straightforward but produce an incomplete picture at the protein level. Proteomics offers physiologically enriched insights to protein function, and ongoing advances are enabling proteome analyses to proceed with increased depth and efficiency. Exciting discoveries have emerged recently amidst growing awareness of the power of proteomics. However, while proven as a potent discovery tool, proteomics is under pressure to provide improved functional value particularly in concert with other investigative approaches. As reviewed here for ERp29, a recently discovered endoplasmic reticulum protein, the role of novel proteins can remain elusive even after substantial information has accrued. Thousands more proteins of uncertain function will be unveiled in the near future. Consequently, the goalposts are moving for proteomics both through increasing demand for high-value functional information and improving capacity to deliver.
Proteomics for cancer biomarker discovery.
Srinivas PR, Verma M, Zhao Y, Srivastava S.
Clin Chem. 2002 Aug;48(8):1160-9.
[ expand abstract ]
The emergence of novel technologies allows researchers to facilitate the comprehensive analyses of genomes, transcriptomes, and proteomes in health and disease. The information that is expected from such technologies may soon exert a dramatic change in the pace of cancer research and impact dramatically on the care of cancer patients. These approaches have already demonstrated the power of molecular medicine in discriminating among disease subtypes that are not recognizable by traditional pathologic criteria and in identifying specific genetic events involved in cancer progression. This review covers a selection of advances in the realm of proteomics and its promise for cancer biomarker discovery. It also addresses issues regarding sample preparation and specificity and discusses current challenges that need to be overcome. Finally, the review touches on the efforts of the Early Detection Research Network at the National Cancer Institute in promoting biomarker discovery for translation at the clinical level.
Is mass spectrometry ready for proteome-wide protein expression analysis?
Rappsilber J, Mann M.
Genome Biol.
2002 Jul 31;3(8):COMMENT2008. Epub 2002 Jul 31.
[ expand abstract ]
Recent advances in mass spectrometry will soon allow routine analysis of protein expression levels. How close are we to true quantitative proteomics?
Molecular biologist's guide to proteomics.
Graves PR, Haystead TA.
Microbiol Mol Biol Rev. 2002 Mar;66(1):39-63; table of contents.
[ expand abstract ]
The emergence of proteomics, the large-scale analysis of proteins, has been inspired by the realization that the final product of a gene is inherently more complex and closer to function than the gene itself. Shortfalls in the ability of bioinformatics to predict both the existence and function of genes have also illustrated the need for protein analysis. Moreover, only through the study of proteins can posttranslational modifications be determined, which can profoundly affect protein function. Proteomics has been enabled by the accumulation of both DNA and protein sequence databases, improvements in mass spectrometry, and the development of computer algorithms for database searching. In this review, we describe why proteomics is important, how it is conducted, and how it can be applied to complement other existing technologies. We conclude that currently, the most practical application of proteomics is the analysis of target proteins as opposed to entire proteomes. This type of proteomics, referred to as functional proteomics, is always driven by a specific biological question. In this way, protein identification and characterization has a meaningful outcome. We discuss some of the advantages of a functional proteomics approach and provide examples of how different methodologies can be utilized to address a wide variety of biological problems.
Organellar proteomics: the prizes and pitfalls of opening the nuclear envelope.
Schirmer EC, Gerace L.
Genome Biol. 2002;3(4):REVIEWS1008.
[ expand abstract ]
Proteomic studies have the potential to comprehensively define the composition of organelles but are limited by the organellar cross-contamination that arises during subcellular fractionation. Comparative proteomics of organellar subfractions can mitigate these problems, as demonstrated by a recent study involving the nuclear envelope.
2001
Proteomics: an holistic analysis of nature's proteins.
Hebestreit HF.
Curr Opin Pharmacol. 2001 Oct;1(5):513-20.
[ expand abstract ]
Proteomics has matured to a technology platform way beyond two-dimensional gel electrophoresis, delivering on its promise to identify structure, function and cellular localization of all proteins expressed in a cell at a given time. Major achievements in the past year include mapping the proteome of human and microbial cells, improvements in two-dimensional gel electrophoresis and mass spectrometric analysis, and the development of protein arrays and biochips.
Proteomics in early detection of cancer.
Srinivas PR, Srivastava S, Hanash S, Wright GL Jr.
Clin Chem. 2001 Oct;47(10):1901-11.
[ expand abstract ]
Early detection is critical in cancer control and prevention. Biomarkers help in this process by providing valuable information about a the status of a cell at any given point in time. As a cell transforms from nondiseased to neoplastic, distinct changes occur that could be potentially detected through the identification of the appropriate biomarkers. Biomarker research has benefited from advances in technology such as proteomics. We discuss here ongoing research in this field, focusing on proteomic technologies. The advances in two-dimensional electrophoresis and mass spectrometry are discussed in light of their contribution to biomarker research. Chip-based techniques, such as surface-enhanced laser desorption, and ionization and emerging methods, such as tissue and antibody arrays, are also discussed. The development of bioinformatic tools that have and are being developed in parallel to proteomics is also addressed. This report brings into focus the efforts of the Early Detection Research Network at the National Cancer Institute in harnessing scientific expertise from leading institutions to identify and validate biomarkers for early detection and risk assessment.
Mass spectrometry in proteomics.
Aebersold R, Goodlett DR.
Chem Rev. 2001 Feb;101(2):269-95.
[ expand abstract ]
(No abstract)
Arrays for protein expression profiling: towards a viable alternative to two-dimensional gel electrophoresis?
Jenkins RE, Pennington SR.
Proteomics.
2001 Jan;1(1):13-29.>
[ expand abstract ]
Two-dimensional gel electrophoresis (2-DE) is used as a platform method for the measurement of protein expression patterns within cells, tissues or organisms. This approach can support expression profiling of several thousand proteins in multiple samples and as such it is currently unrivaled as a tool for the analysis of protein expression, which is a key component of the rapidly expanding field of proteomics. However, 2-DE has a number of significant limitations and as a consequence, alternative approaches for the measurement of expression of proteins within complex samples are actively being explored. Here we review some existing and emerging methods for protein expression analysis. In particular, we review a range of technologies that might be integrated to support the development of 'arrays' or 'chips' for rapid, high-throughput analysis of protein expression in a manner analogous to the current use of DNA arrays for mRNA expression analysis. We conclude that such separation-independent platforms may ultimately supersede two-dimensional (2-D) gel-based analyses for global protein expression analysis but that before this the technologies might provide important new platforms for diagnostic and prognostic monitoring of diseases.
Functional proteomics: large-scale analysis of protein kinase activity.
Lawrence DS.
Genome Biol. 2001;2(2):REVIEWS1007.
[ expand abstract ]
Proteome-wide sampling of function can be used to shed light on complex biological systems. Protein microarrays have now been used to investigate the substrate specificities of essentially all the protein kinases encoded by the yeast genome.
2000
Whole genomes: the foundation of new biology and medicine.
Broder S, Venter JC.
Curr Opin Biotechnol. 2000 Dec;11(6):581-5.
[ expand abstract ]
Our genomic DNA sequence provides a unique glimpse of the provenance and evolution of our species, the migration of peoples, and the causation of disease. Understanding the genome may help resolve previously unanswerable questions, including perhaps which human characteristics are innate or acquired. Such an understanding will make it possible to study how genomic DNA sequence varies among populations and among individuals, including the role of such variation in the pathogenesis of important illnesses and responses to pharmaceuticals. The study of the genome and the associated proteomics of free-living organisms will eventually make it possible to localize and annotate every human gene, as well as the regulatory elements that control the timing, organ-site specificity, extent of gene expression, protein levels, and post-translational modifications. For any given physiological process, we will have a new paradigm for addressing its evolution, development, function, and mechanism.
Proteomics: new perspectives, new biomedical opportunities.
Banks RE, Dunn MJ, Hochstrasser DF, Sanchez JC, Blackstock W, Pappin DJ, Selby PJ.
Lancet. 2000 Nov 18;356(9243):1749-56.
[ expand abstract ]
Proteomics-based approaches, which examine the expressed proteins of a tissue or cell type, complement the genome initiatives and are increasingly being used to address biomedical questions. Proteins are the main functional output, and the genetic code cannot always indicate which proteins are expressed, in what quantity, and in what form. For example, post-translational modifications of proteins, such as phosphorylation or glycosylation, are very important in determining protein function. Similarly, the effects of environmental factors or multigenic processes such as ageing or disease cannot be assessed simply by examination of the genome alone. This review describes the underlying technology and illustrates several areas of biomedical research, ranging from pathogenesis of neurological disorders to drug and vaccine design, in which potential clinical applications are being explored.
The use of recombinant antibodies in proteomics.
Holt LJ, Enever C, de Wildt RM, Tomlinson IM.
Curr Opin Biotechnol. 2000 Oct;11(5):445-9.
[ expand abstract ]
Recombinant antibodies are becoming increasingly important in the field of proteomics. Recent advances include the development of large phage-antibody libraries that contain high-affinity binders to almost any target protein, and new methods for high-throughput selection of antibody-antigen interactions. Coupled with a range of new screening technologies that use high-density antibody arrays to identify differentially expressed proteins, these antibody libraries can be applied to whole proteome analysis
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