Author: Huili Zhai
Publisher:
ISBN:
Category :
Languages : en
Pages : 332

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Book Description

Author: Julian Whitelegge
Publisher: Elsevier
ISBN: 0080932037
Category : Science
Languages : en
Pages : 563

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Book Description
This book is designed to be a central text for young graduate students interested in mass spectrometry as it relates to the study of protein structure and function as well as proteomics. It is a definite must-have work for:- libraries at academic institutions with Master and Graduate programs in biochemistry, molecular biology, structural biology and proteomics- individual laboratories with interests covering these areas - libraries and individual laboratories in the pharmaceutical and biotechnology industries.*Serves as an essential reference to those working in the field*Incorporates the contributions of prominent experts *Features comprehensive coverage and a logical structure

Author: Yutong Jin
Publisher:
ISBN:
Category :
Languages : en
Pages : 0

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Book Description
Mass spectrometry (MS)-based proteomics, including top-down, middle-down and bottom-up approaches, has become the method of choice for protein identification, quantification, and characterization. Top-down proteomics represents a superior approach for comprehensive protein characterization by analyzing intact proteins, providing an overview of all the proteoforms present in a sample and allowing for identification of various proteoforms and in-depth characterization of sequences and post-translational modifications (PTMs). However, the protein mass range that can be analyzed by top-down approach is normally limited to 100 kDa. Middle-down proteomics, analyzing polypeptides from proteolysis, is a complementary strategy to top-down proteomics for characterization of large proteins (>100 kDa) and complex PTMs. In this dissertation, I developed and applied top-down and middle-down proteomics to comprehensively characterize the sarcomeric proteins from striated muscle tissues. Sarcomeric proteins are expressed in different isoforms produced by homologous genes, in the presence of various proteoforms produced by a single gene via genetic variations, alternative RNA splicing, and PTMs. The various isoforms and PTMs of sarcomeric proteins are associated with muscle contractile properties and thus affects the entire muscle function. However, a comprehensive characterization of sarcomeric protein isoforms and PTMs is still challenging due to the high complexity of the muscle proteome and the limitations in techniques for protein separation and characterization. The first part of this dissertation focuses on characterizing sarcomeric proteins below 100 kDa using top-down proteomics. Sarcomeric proteins from multiple skeletal muscle tissues were characterized by liquid chromatography and mass spectrometry plus (LC-MS+) strategy which combines online LC-MS and offline tandem MS (MS/MS) analysis. I also developed a reversed-phase chromatography method using monolithic column for highly efficient separation of sarcomeric proteins and combined it with high-resolution MS for top-down proteomics. The second part of this dissertation describes the development of middle-down proteomics for characterization of large sarcomeric proteins (>100 kDa). I developed a MS-compatible size-exclusion chromatography (SEC) method to purify myosin heavy chain from cardiac muscle tissue and characterize this protein by refining a middle-down MS strategy. I also developed a middle-down proteomics strategy to study the phosphorylation status of a serine-arginine-rich protein RNA-binding motif 20. Finally, combining top-down and middle-down proteomics, I described a strategy for the comprehensive characterization of a monoclonal antibody. Collectively, this dissertation provides comprehensive analysis of the isoforms and PTMs of sarcomeric proteins that will help elucidate molecular event in muscle contraction and disease pathologies. Furthermore, this dissertation presents a variety of novel LC and MS methods for protein separation and characterization, enabling the further characterization of other proteomes with deep proteome coverage.

Author: David D. Weis
Publisher: John Wiley & Sons
ISBN: 1118616499
Category : Science
Languages : en
Pages : 422

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Book Description
Hydrogen exchange mass spectrometry is widely recognized for its ability to probe the structure and dynamics of proteins. The application of this technique is becoming widespread due to its versatility for providing structural information about challenging biological macromolecules such as antibodies, flexible proteins and glycoproteins. Although the technique has been around for 25 years, this is the first definitive book devoted entirely to the topic. Hydrogen Exchange Mass Spectrometry of Proteins: Fundamentals, Methods and Applications brings into one comprehensive volume the theory, instrumentation and applications of Hydrogen Exchange Mass Spectrometry (HX-MS) - a technique relevant to bioanalytical chemistry, protein science and pharmaceuticals. The book provides a solid foundation in the basics of the technique and data interpretation to inform readers of current research in the method, and provides illustrative examples of its use in bio- and pharmaceutical chemistry and biophysics In-depth chapters on the fundamental theory of hydrogen exchange, and tutorial chapters on measurement and data analysis provide the essential background for those ready to adopt HX-MS. Expert users may advance their current understanding through chapters on methods including membrane protein analysis, alternative proteases, millisecond hydrogen exchange, top-down mass spectrometry, histidine exchange and method validation. All readers can explore the diversity of HX-MS applications in areas such as ligand binding, membrane proteins, drug discovery, therapeutic protein formulation, biocomparability, and intrinsically disordered proteins.

Author: W. Andy Tao
Publisher: John Wiley & Sons
ISBN: 1118970217
Category : Science
Languages : en
Pages : 448

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Book Description
PROVIDES STRATEGIES AND CONCEPTS FOR UNDERSTANDING CHEMICAL PROTEOMICS, AND ANALYZING PROTEIN FUNCTIONS, MODIFICATIONS, AND INTERACTIONS—EMPHASIZING MASS SPECTROMETRY THROUGHOUT Covering mass spectrometry for chemical proteomics, this book helps readers understand analytical strategies behind protein functions, their modifications and interactions, and applications in drug discovery. It provides a basic overview and presents concepts in chemical proteomics through three angles: Strategies, Technical Advances, and Applications. Chapters cover those many technical advances and applications in drug discovery, from target identification to validation and potential treatments. The first section of Mass Spectrometry-Based Chemical Proteomics starts by reviewing basic methods and recent advances in mass spectrometry for proteomics, including shotgun proteomics, quantitative proteomics, and data analyses. The next section covers a variety of techniques and strategies coupling chemical probes to MS-based proteomics to provide functional insights into the proteome. In the last section, it focuses on using chemical strategies to study protein post-translational modifications and high-order structures. Summarizes chemical proteomics, up-to-date concepts, analysis, and target validation Covers fundamentals and strategies, including the profiling of enzyme activities and protein-drug interactions Explains technical advances in the field and describes on shotgun proteomics, quantitative proteomics, and corresponding methods of software and database usage for proteomics Includes a wide variety of applications in drug discovery, from kinase inhibitors and intracellular drug targets to the chemoproteomics analysis of natural products Addresses an important tool in small molecule drug discovery, appealing to both academia and the pharmaceutical industry Mass Spectrometry-Based Chemical Proteomics is an excellent source of information for readers in both academia and industry in a variety of fields, including pharmaceutical sciences, drug discovery, molecular biology, bioinformatics, and analytical sciences.

Author: Roza Maria Kamp
Publisher: Springer Science & Business Media
ISBN: 3642592198
Category : Science
Languages : en
Pages : 311

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Book Description
"Protein Structure Analysis - Preparation and Characterization" is a compilation of practical approaches to the structural analysis of proteins and peptides. Here, about 20 authors describe and comment on techniques for sensitive protein purification and analysis. These methods are used worldwide in biochemical and biotechnical research currently being carried out in pharmaceu tical and biomedical laboratories or protein sequencing facilities. The chapters have been written by scientists with extensive ex perience in these fields, and the practical parts are well documen ted so that the reader should be able to easily reproduce the described techniques. The methods compiled in this book were demonstrated in student courses and in the EMBO Practical Course on "Microsequence Analysis of Proteins" held in Berlin September 10-15, 1995. The topics also derived from a FEBS Workshop, held in Halkidiki, Thessaloniki, Greece, in April, 1995. Most of the authors participated in these courses as lecturers and tutors and made these courses extremely lively and successful. Since polypeptides greatly vary depending on their specific structure and function, strategies for their structural analysis must for the most part be adapted to each individual protein. Therefore, advantages and limitations of the experimen tal approaches are discussed here critically, so that the reader becomes familiar with problems that might be encountered.

Author: Robert A. Copeland
Publisher: John Wiley & Sons
ISBN: 0471723266
Category : Science
Languages : en
Pages : 295

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Book Description
Vital information for discovering and optimizing new drugs "Understanding the data and the experimental details that support it has always been at the heart of good science and the assumption challenging process that leads from good science to drug discovery. This book helps medicinal chemists and pharmacologists to do exactly that in the realm of enzyme inhibitors." -Paul S. Anderson, PhD This publication provides readers with a thorough understanding of enzyme-inhibitor evaluation to assist them in their efforts to discover and optimize novel drug therapies. Key topics such as competitive, noncompetitive, and uncompetitive inhibition, slow binding, tight binding, and the use of Hill coefficients to study reaction stoichiometry are all presented. Examples of key concepts are presented with an emphasis on clinical relevance and practical applications. Targeted to medicinal chemists and pharmacologists, Evaluation of Enzyme Inhibitors in Drug Discovery focuses on the questions that they need to address: * What opportunities for inhibitor interactions with enzyme targets arise from consideration of the catalytic reaction mechanism? * How are inhibitors evaluated for potency, selectivity, and mode of action? * What are the advantages and disadvantages of specific inhibition modalities with respect to efficacy in vivo? * What information do medicinal chemists and pharmacologists need from their biochemistry and enzymology colleagues to effectively pursue lead optimization? Beginning with a discussion of the advantages of enzymes as targets for drug discovery, the publication then explores the reaction mechanisms of enzyme catalysis and the types of interactions that can occur between enzymes and inhibitory molecules that lend themselves to therapeutic use. Next are discussions of mechanistic issues that must be considered when designing enzyme assays for compound library screening and for lead optimization efforts. Finally, the publication delves into special forms of inhibition that are commonly encountered in drug discovery efforts, but can be easily overlooked or misinterpreted. This publication is designed to provide students with a solid foundation in enzymology and its role in drug discovery. Medicinal chemists and pharmacologists can refer to individual chapters as specific issues arise during the course of their ongoing drug discovery efforts.

Author:
Publisher:
ISBN:
Category : Dissertations, Academic
Languages : en
Pages : 858

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Book Description

Author: John R. Griffiths
Publisher: John Wiley & Sons
ISBN: 1119045851
Category : Science
Languages : en
Pages : 414

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Book Description
Covers all major modifications, including phosphorylation, glycosylation, acetylation, ubiquitination, sulfonation and and glycation Discussion of the chemistry behind each modification, along with key methods and references Contributions from some of the leading researchers in the field A valuable reference source for all laboratories undertaking proteomics, mass spectrometry and post-translational modification research

Author: Jozef Samaj
Publisher: Springer Science & Business Media
ISBN: 3540726179
Category : Science
Languages : en
Pages : 397

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Book Description
Plant Proteomics highlights rapid progress in this field, with emphasis on recent work in model plant species, sub-cellular organelles, and specific aspects of the plant life cycle such as signaling, reproduction and stress physiology. Several chapters present a detailed look at diverse integrated approaches, including advanced proteomic techniques combined with functional genomics, bioinformatics, metabolomics and molecular cell biology, making this book a valuable resource for a broad spectrum of readers.