Manual of Cardiovascular Proteomics


Book Description

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.




Cardiovascular Proteomics


Book Description

This cutting-edge book presents protocols and strategies for proteomic evaluation of cardiovascular disease written by pioneering researchers in the field. Topics explored in this comprehensive volume include obtaining specific heart proteins, techniques for identifying risk biomarkers of atherome plaque rupture, analyzing the secretome of explanted endarterectomies cultured in vitro, and phage display techniques for deciphering the molecular diversity of blood vessels.




Heart Proteomics


Book Description

The use of proteomics to study complex diseases such as cardiovascular disease, the leading cause of death in developed countries, has grown exponentially in recent years. Proteomics is a rapidly expanding investigation platform in cardiovascular medicine and is becoming integrated and incorporated into cardiovascular research. The proteomics field continues to develop with major improvements in mass spectrometry instrumentation, methodology and data analysis. Heart Proteomics: Methods and Protocols complies a selection of techniques and methods that target the numerous processes implicated in the pathophysiology of heart. Chapters cover protocols and updated methods in the heart proteomic area with a particular focus on MS-based methods of protein and peptide quantification and the analysis of posttranslational modifications as well as descriptions of system biology approaches, which provide a better understanding of normal and pathological processes. Written in the successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Heart Proteomics: Methods and Protocols is a representative selection of methods that will prove to be a useful resource for experienced proteomics practitioners and newcomers alike.




Proteomic and Genomic Analysis of Cardiovascular Disease


Book Description

This is the very first book to focus on this new approach that will eventually aid in developing new diagnostic markers and therapies for controlling and treating heart disease - the number-one killer in the industrialized world. Divided into two parts, the book describes not only the potentials, but also the limitations of these technologies. The editors, both well known within the scientific community, provide new insights into the biochemical and cellular mechanisms of cardiovascular disease, as well as covering the transition into clinical applications. In so doing, they highlight the various strategies and technical aspects so as to assist the growing number of researchers intending to utilize these approaches. The result is an excellent way of educating and informing graduate students, post-doctoral fellows as well as researchers in academia and industry about the latest developments in this area.




Cardiovascular Proteomics


Book Description

"Cardiovascular Proteomics: Methods and Protocols" covers many of the aspects of the proteomic approach in the cardiovascular field. This volume takes the reader through the complete process of proteomic analysis, from the obtention of specific heart proteins (troponin I) to the new techniques of identifying risk biomarkers of atherome plaque rupture analyzing the secretome of explanted endartrectomies cultured in vitro or the application of phage display techniques to decipher the molecular diversity of blood vessels.




Proteomics in Drug Research


Book Description

From skillful handling of the wide range of technologies to successful applications in drug discovery -- this handbook has all the information professional proteomics users need. Edited by experts working at one of the hot spots in European proteomic research, the numerous contributions by experts from the pharmaceutical industry and public proteomics consortia to provide the necessary perspective on current trends and developments in this exciting field. Following an introductory chapter, the book moves on to proteomic technologies, such as protein biochips, protein-protein interactions, and proteome analysis in situ. The section on applications includes bioinformatics, Alzheimer's disease, neuroproteomics, plasma and T-cell proteomics, differential phosphoproteome analysis and biomarkers, as well as pharmacogenomics. Invaluable reading for medicinal and pharmaceutical chemists, gene technologists, molecular biologists, and those working in the pharmaceutical industry.




Defining the Human Heart Proteoform Landscape with Top-Down Proteomics


Book Description

Functional diversity in the heart proteome is attributed to the vast array of proteoforms arising from molecular processing events such as alternative splicing and post-translational modifications (PTMs). Given the importance of isoform switching and PTMs in the regulation of heart function and dysfunction, it is important to study the heart proteome at the proteoform level in our effort to understand heart health and disease. Top-down proteomics featuring high-resolution mass spectrometry (MS) is a premier tool for studying proteoforms in biological systems. However, the high complexity and wide dynamic range of the heart proteome have precluded in-depth top-down proteomic analysis using conventional platforms. Moreover, studying large proteoforms (>60 kDa) with top-down proteomics is especially challenging due to reduced MS signal-to-noise ratio with increasing size combined with signal suppression caused by the co-elution of smaller, more abundant proteoforms. To improve coverage of the heart proteoform landscape and access large proteoforms by top-down proteomics, serial size-exclusion chromatography (sSEC) was introduced for size-based fractionation of complex protein mixtures. A two-dimensional separation platform coupling sSEC with reversed-phase chromatography and high-resolution MS expanded coverage of heart proteoforms and enabled the detection of those up to 223 kDa from heart tissue lysate (Chapter 3). Despite improved MS detection of large proteoforms, challenges associated with tandem MS (MS/MS) fragmentation and limitations in mass resolution still prevented their identification. Intact-mass analysis software relating experimental proteoform mass to a list of candidate proteoforms enabled interpretation of complex top-down proteomic data and facilitated the identification of large heart proteoforms, including the 140-kDa myosin binding protein C (Chapter 4). Fourier transform ion cyclotron resonance (FT-ICR) MS has also provided a versatile platform for characterization of proteoforms, given its high mass resolving power and mass accuracy combined with multi-faceted fragmentation capabilities (Chapter 5). sSEC fractionation paired with top-down FT-ICR MS allowed for straightforward characterization of metabolic enzymes extracted from heart tissue (Chapter 6). Finally, top-down proteomics was applied to investigate the genotype-proteoform phenotype relationship in human hypertrophic cardiomyopathy (HCM), the most common heritable heart disease (Chapter 7). We quantified contractile proteoforms in septal myectomy tissues from HCM patients and observed converging proteoform phenotypes, regardless of disease-causing mutation, suggesting that common pathways are activated during disease progression and underscoring the importance of proteoform-level analysis. Overall, the studies detailed in this dissertation have charted new territory in the heart proteoform landscape, especially for heart proteoforms with molecular weight above 60 kDa. sSEC-based top-down proteomics facilitated cataloging of heart proteoforms to aid future studies seeking to investigate proteoforms in heart health and disease (Chapter 8). Future applications of the technology detailed herein will expand on SEC-based techniques for top-down proteomics and adapt the technology to accommodate the study of protein interactomes and large proteoforms from minute samples, such as those obtained from patients with heart disease.




Mass Spectrometry-Based Chemical Proteomics


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.




Platelet Proteomics


Book Description

The purpose of the book is to introduce platelets, and their functional role in thrombotic and cardiovascular disease, justifying the relevance of platelet proteomics research. Focus then shifts to the recent developments on mass spectrometry (MS)-based proteomics. This chapter shows potential applications for platelet proteomics not yet carried out. It includes examples of post-translational modifications (PTMs) analysis in platelets. The second part of the book focuses on the main research done so far on platelet proteomics. This includes general proteome mapping by non-gel based separation methods (MudPit), analysis of the general platelet proteome and signaling cascades by gel-based separation methods (2-DE), sub-proteome analyses (secretome/releasate, membrane proteins, organelles). Finally, the last section links the platelet transcriptome and application to disease. This section is highly relevant and includes chapters on proteomics, transcriptomics, functional genomics, systems biology, and their applications to platelet-related diseases.




Proteomics


Book Description

Biomedical research has entered a new era of characterizing a disease or a protein on a global scale. In the post-genomic era, Proteomics now plays an increasingly important role in dissecting molecular functions of proteins and discovering biomarkers in human diseases. Mass spectrometry, two-dimensional gel electrophoresis, and high-density antibody and protein arrays are some of the most commonly used methods in the Proteomics field. This book covers four important and diverse areas of current proteomic research: Proteomic Discovery of Disease Biomarkers, Proteomic Analysis of Protein Functions, Proteomic Approaches to Dissecting Disease Processes, and Organelles and Secretome Proteomics. We believe that clinicians, students and laboratory researchers who are interested in Proteomics and its applications in the biomedical field will find this book useful and enlightening. The use of proteomic methods in studying proteins in various human diseases has become an essential part of biomedical research.