Shotgun Proteomics

Shotgun Proteomics: Methods and Protocols serves as a vital collection of protocols through which thousands of proteins can be simultaneously identified, quantified, and characterized in a high throughput manner. Beginning with the history of proteomics centered on the vital role of mass spectrometry in its development, this detailed volume continues with chapters on sample pre-fractionation, in vivo and in vitro stable isotope labeling, label-free proteomics, informatics, protein-protein interactions, targeted proteomics, and post-translational modifications. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Practical and comprehensive, Shotgun Proteomics: Methods and Protocols is an ideal and up-to-date guide for researchers seeking to understand the proteome of any given species.

In this thesis, the author outlines the development of new monolithic columns and isotope dimethyl labeling strategies and their applications in high-performance proteome analyses. Though different types of monolithic columns have been widely developed for chromatography and electrophoresis separation, their application in proteomics for complex peptide mixtures separation is still a challenge. The author discusses the preparation of new monolithic columns and optimization of chromatography separation capability to improve coverage and accuracy of proteome analysis. Further, the author describes a novel online multidimensional chromatography system combined with automated online isotope labeling, which significantly improves the throughput, sensitivity and accuracy of quantitative proteomics. In addition to the development of new technologies, the author investigates the proteome and phosphoproteome expression changes of clinical hepatocellular carcinoma tissues and the hippocampi of mice with Alzheimer’s disease. The work in this thesis has led to several publications in high-profile journals in the fields of analytical chemistry and proteome research.

Facilitating the innovation, development, and application of new spectroscopic methods in proteomics, Spectral Techniques in Proteomics provides a broad overview of the spectroscopic toolbox that can be used, either with proteome or sub-proteome mixtures or with individual/purified proteins studied in parallel. It gives a modest overview of

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.

This volume is the first collection of applications of proteomics to analyze various human body fluids. Proteomics of Human Bodyfluids consists of two parts. The first provides basic principles and strategies for proteomic analysis of human body fluids. The second offers more details regarding methodologies and recent findings and clinical applications of each specific type of human body fluids.

Proteomics: Methods Express identifies the most powerful new technologies and presents them in a way that allows their robust implementation. The focus is on proteomic methods and strategies that are reliable and of general applicability. Each chapter presents descriptions of what can, and cannot, be achieved with the relevant procedures so that readers can make informed judgments prior to establishing the methods in-house. Every chapter discusses the merits and limitations of various approaches then provides tried-and-tested protocols with hints and tips for success and troubleshooting for when things go wrong.

This is one of the first books to focus on the dynamic aspect of proteomes. The book introduces proteomics to the newcomer, reviews the theoretical aspects of proteomics and its state-of-the art technologies, along with a number of biological applications using "classical" proteomic technology. The book also presents a new concept, the Dynamome, or the expression of a comprehensive molecular set that participates in the whole dynamic process of a series of cellular events.

This book fulfils the need to keep up with the high number of innovations in proteomics, and at the same time to warn the readers about the danger of manufacturers and scientists claims around new technologies. Mass spectrometry stands as the core technology in proteomics. The emerging field of targeted proteomics and its potential applications in the cardiovascular arena are also reviewed and discussed. A concluding section highilghts the promise of proteomics in the light of these recent developments.​ As this technique and its applications have undergone remarkable advances in the past years, recent updates on proteomic applications are covered. Another key concept revealed by proteomic technologies is that the extent of protein post-translational modifications (PTMs) as well as their impact on the phenotype has been underestimated by pre-proteomics science. As such, part of the manual focuses on the emerging role of PTMs in basic cardiovascular sciences and in the clinics. In fact, there is an emerging consensus that the detailed annotation of protein PTMs could lead to a more in-depth representation of biological systems, translating into more specific targets for therapy as well as biomarkers. Moreover, a recent trend is so-called “targeted proteomics”. The approach was awarded the title of “Method of the Year” by Nature in 2013 (see the editorial by Vivien Marx in 1st issue of Nature in Jan 2013). According to a few proteomic scientists the emphasis should not be placed on generating long lists of proteins but lists of proteins with a true biological meaning.