This volume supplements Volumes 63, 64, 87, and 249 of Methods in Enzymology. These volumes provide a basic source for the quantitative interpretation of enzyme rate data and the analysis of enzyme catalysis. Among the major topics covered are Engergetic Coupling in Enzymatic Reactions, Intermediates and Complexes in Catalysis, Detection and Properties of Low Barrier Hydrogen Bonds, Transition State Determination, and Inhibitors.The critically acclaimed laboratory standard for more than forty years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with more than 300 volumes (all of them still in print), the series contains much material still relevant today--truly an essential publication for researchers in all fields of life sciences.

Fully updated and expanded-a solid foundation for understandingexperimental enzymology. This practical, up-to-date survey is designed for a broadspectrum of biological and chemical scientists who are beginning todelve into modern enzymology. Enzymes, Second Editionexplains the structural complexities of proteins and enzymes andthe mechanisms by which enzymes perform their catalytic functions.The book provides illustrative examples from the contemporaryliterature to guide the reader through concepts and data analysisprocedures. Clear, well-written descriptions simplify the complexmathematical treatment of enzyme kinetic data, and numerouscitations at the end of each chapter enable the reader to accessthe primary literature and more in-depth treatments of specifictopics. This Second Edition of Enzymes: A Practical Introductionto Structure, Mechanism, and Data Analysis features refinedand expanded coverage of many concepts, while retaining theintroductory nature of the book. Important new featuresinclude: A new chapter on protein-ligand binding equilibria Expanded coverage of chemical mechanisms in enzyme catalysisand experimental measurements of enzyme activity Updated and refined discussions of enzyme inhibitors andmultiple substrate reactions Coverage of current practical applications to the study ofenzymology Supplemented with appendices providing contact information forsuppliers of reagents and equipment for enzyme studies, as well asa survey of useful Internet sites and computer software forenzymatic data analysis, Enzymes, Second Edition isthe ultimate practical guide for scientists and students inbiochemical, pharmaceutical, biotechnical, medicinal, andagricultural/food-related research.

This volume in the Advances in Protein Chemistry series features cutting-edge articles on topics in protein chemistry. This volume includes chapters on the structural basis of effector regulation and signal termination in heterotrimeric Gƒуnƒnproteins; How do receptors activate G proteins; Some mechanistic insights into GPCR activation from detergent solubilized ternary complexes on beads; Activation of G protein coupled receptors; Kinetic analysis of g-protein-coupled receptor signaling using fluorescence resonance energy transfer in living cells; Regulation of Rho Guanine Nucleotide Exchange Factors (RhoGEFs) by G proteins.

Specific complexes of protein and RNA carry out many essential biological functions, including RNA processing, RNA turnover, RNA folding, as well as the translation of genetic information from mRNA into protein sequences. Messenger RNA (mRNA) decay is now emerging as an important control point and a major contributor to gene expression. Continuing identification of the protein factors and cofactors, and mRNA instability elements responsible for mRNA decay allow researchers to build a comprehensive picture of the highly orchestrated processes involved in mRNA decay and its regulation. - Covers the nonsense-mediated mRNA decay (NMD) or mRNA surveillance pathway - Expert researchers introduce the most advanced technologies and techniques to identify mRNA processing, transport, localization and turnover, which are central to the process of gene expression - Offers step-by-step lab instructions, including necessary equipment and reagents

Systems biology is a term used to describe a number of trends in bioscience research and a movement that draws on those trends. This volume in the Methods in Enzymology series comprehensively covers the methods in systems biology. With an international board of authors, this volume is split into sections that cover subjects such as machines for systems biology, protein production and quantification for systems biology, and enzymatic assays in systems biology research. - This volume in the Methods in Enzymology series comprehensively covers the methods in systems biology - With an international board of authors, this volume is split into sections that cover subjects such as machines for systems biology, protein production and quantification for systems biology, and enzymatic assays in systems biology research

There are numerous excellent reviews on fragment-based drug discovery (FBDD), but there are to date no hand-holding guides or protocols with which one can embark on this orthogonal approach to complement traditional high throughput screening methodologies. This Methods in Enzymology volume offers the tools, practical approaches, and hit-to-lead examples on how to conduct FBDD screens. The chapters in this volume cover methods that have proven to be successful in generating leads from fragments, including chapters on how to apply computational techniques, nuclear magnetic resonance, surface plasma resonance, thermal shift and binding assays, protein crystallography, and medicinal chemistry in FBDD. Also elaborated by experienced researchers in FBDD are sample preparations of fragments, proteins, and GPCR as well as examples of how to generate leads from hits. - Offers the tools, practical approaches, and hit-to-lead examples on how to conduct FBDD screens - The chapters in this volume cover methods that have proven to be successful in generating leads from fragments, including chapters on how to apply computational techniques, nuclear magnetic resonance, surface plasma resonance, thermal shift and binding assays, protein crystallography, and medicinal chemistry in FBDD

In this 3 volume collection focusing on glycomics, readers will appreciate how such discoveries were made and how such methods can be applied for readers' own research efforts - Each chapter has been designed so that enough scientific background will be given in each chapter for further development of methods by readers themselves - Useful for all levels of scientists starting from the last years of colleges, graduate students, postdoctoral fellows to professors and to all levels of scientists in research institutes including industry

Guide to Yeast Genetics and Molecular Biology presents, for the first time, a comprehensive compilation of the protocols and procedures that have made Saccharomyces cerevisiae such a facile system for all researchers in molecular and cell biology. Whether you are an established yeast biologist or a newcomer to the field, this volume contains all the up-to-date methods you will need to study "Your Favorite Gene" in yeast.Key Features* Basic Methods in Yeast Genetics* Physical and genetic mapping* Making and recovering mutants* Cloning and Recombinant DNA Methods* High-efficiency transformation* Preparation of yeast artificial chromosome vectors* Basic Methods of Cell Biology* Immunomicroscopy* Protein targeting assays* Biochemistry of Gene Expression* Vectors for regulated expression* Isolation of labeled and unlabeled DNA, RNA, and protein

This volume of Methods in Enzymology looks at Gene Transfer Vectors for Clinical Application. The chapters provide an invaluable resource for academics, researchers and students alike. With an international board of authors, this volume covers such topics as General principles of retrovirus vector design, Chronic granulomatous disease (CGD), Gene therapy for blindness, and Retrovirus genetic strategy and vector design. Chapters provide an invaluable resource for academics, researchers and students alike International board of authors This volume covers such topics as general principles of retrovirus vector design, chronic granulomatous disease (CGD), gene therapy for blindness, and retrovirus genetic strategy and vector design