Protein Lipidation Protocols
Protein Lipidation Protocols

Author: Michael H. Gelb

Publisher: Springer Science & Business Media

Published: 2008-02-03

Total Pages: 243

ISBN-13: 1592592643

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In Protein Lipidation Protocols, Michael Gelb brings together a collection of readily reproducible techniques for studying protein lipidation, the covalent attachment of lipids to proteins. These cutting-edge methods-many never published before in a "hands-on" format-deal with glycosyl phosphatidylinositol (GPI)-containing compounds, protein fatty acylation, and protein prenylation. Included are novel techniques for determining the chemical structure of GPI-anchors, for radiolabeling the prenyl groups of protein in eukaryotic cells, a tool for developing inhibitors of the protein farnesyltransferase, and for an exciting lysosomal enzyme that cleaves fatty acyl groups from proteins, the first fatty acylase discovered. Protein Lipidation Protocols offers biochemists, cell and molecular biologists, medicinal chemists, and pharmaceutical researchers state-of-the-art tools for understanding the complex biochemistry of protein lipidation, as well as catalyzing the development of many important new biopharmaceuticals, including anticancer drugs.

Protein Lipidation
Protein Lipidation

Author: Maurine E. Linder

Publisher: Humana

Published: 2019-06-01

Total Pages: 327

ISBN-13: 9781493995318

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This volume explores techniques used to detect lipids attached to proteins, to analyze the function of lipid modifications, and to characterize the enzymes that add and remove lipids from proteins. The book is organized into seven parts: Part One describes chemically-based strategies to identify substrates for protein lipidation that can be applied to individual proteins or globally using proteomics. Part Two focuses on the enzymes that remove fatty acids from proteins and provides methods to monitor protein biogenesis and palmitate turnover. Part Three addresses biochemical and cellular characterization of DHHC S-acyltransferases, a family of enzymes with 23 members encoded by the human genome. Part Four presents the SwissPalm 2 database and tips on how to use it effectively. Part Five focuses on fatty acylation that occurs in the lumen of the secretory pathway. Parts Six and Seven conclude the book with methods to produce and assay lipid-modified and integral membrane proteins. Written in the highly successful Methods in Molecular Biology series format, 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. Cutting-edge and authoritative, Protein Lipidation: Methods and Protocols is a valuable resource for experts in the field and for investigators who encounter protein lipidation through their research on a particular cellular process or favorite protein.

Calpain Methods and Protocols
Calpain Methods and Protocols

Author: John S. Elce

Publisher: Springer Science & Business Media

Published: 2008-02-05

Total Pages: 340

ISBN-13: 1592590500

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The purpose of Calpain Methods and Protocols is quite straightf- ward: it is to present the actual experimental methods used in many different laboratories for the study of calpain. It will provide the vital experimental detail, and the discussion of possible pitfalls, for which the standard journals no longer provide space. This will make it as easy as possible for investi- tors interested in calpain to adopt established methods without repeating old mistakes, and to adapt and apply these methods in novel approaches to the many outstanding calpain questions. These questions range from purely biochemical problems of protein structure and enzyme regulation at the molecular level, through large areas of cell biology, to applied and clinical aspects of calpain function in human d- ease. Within this panoply of topics, a wide range of investigators will find many fascinating and as yet unanswered questions about calpain. Calpain Methods and Protocols will provide instant access to many essential te- niques, while saving them the time and effort involved in developing a new method. In addition to questions relating to the normal physiological roles of the calpains, there is considerable evidence that inappropriate calpain activity may have pathological effects in many tissues, for example, following ischemia. This provides a major stimulus for the development of specific calpain inhi- tors for therapeutic purposes, and for the development of methods to evaluate such inhibitors.

Bioinformatics Methods and Protocols
Bioinformatics Methods and Protocols

Author: Stephen Misener

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 495

ISBN-13: 1592591922

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Computers have become an essential component of modern biology. They help to manage the vast and increasing amount of biological data and continue to play an integral role in the discovery of new biological relationships. This in silico approach to biology has helped to reshape the modern biological sciences. With the biological revolution now among us, it is imperative that each scientist develop and hone today’s bioinformatics skills, if only at a rudimentary level. Bioinformatics Methods and Protocols was conceived as part of the Methods in Molecular Biology series to meet this challenge and to provide the experienced user with useful tips and an up-to-date overview of current developments. It builds upon the foundation that was provided in the two-volume set published in 1994 entitled Computer Analysis of Sequence Data. We divided Bioinformatics Methods and Protocols into five parts, including a thorough survey of the basic sequence analysis software packages that are available at most institutions, as well as the design and implemen- tion of an essential introductory Bioinformatics course. In addition, we included sections describing specialized noncommercial software, databases, and other resources available as part of the World Wide Web and a stimul- ing discussion of some of the computational challenges biologists now face and likely future solutions.

Integrin Protocols
Integrin Protocols

Author: Anthony R. Howlett

Publisher: Springer Science & Business Media

Published: 2008-02-03

Total Pages: 296

ISBN-13: 159259249X

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In Integrin Protocols, Anthony Howlett and a distinguished panel of experimentalists describe in detail a series of cutting-edge methods for dissecting the role of integrins in biological processes. This wide-ranging collection includes protocols for the analysis of integrin expression-at both the RNA and protein levels-and for elucidating the functional properties of integrins, including those at the cellular level. Each method provides step-by-step instructions for easy reproducibility, along with extensive notes about potential pitfalls, and tips on how to avoid failure. The emphasis is always on the practical steps necessary for experimental success and robust results. Offering powerful tools for understanding how integrins regulate cell growth, differentiation, migration, invasion, angiogenesis, and apoptosis, as well as how abnormalities of integrin expression and function may be implicated in various pathologic conditions, Integrin Protocols constitutes a gold-standard collection of techniques for both new and experienced investigators of the molecular and cellular basis of cardiovascular disease, inflammatory disorders, and cancer.

Connexin Methods and Protocols
Connexin Methods and Protocols

Author: Roberto Bruzzone

Publisher: Springer Science & Business Media

Published: 2008-02-05

Total Pages: 495

ISBN-13: 1592590438

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Direct cell–cell communication is a common property of multicellular organisms that is achieved through membrane channels which are organized in gap junctions. The protein subunits of these intercellular channels, the connexins, form a multigene family that has been investigated in great detail in recent years. It has now become clear that, in different tissues, connexins speak several languages that control specific cellular functions. This progress has been made possible by the availability of new molecular tools and the improvement of basic techniques for the study of membrane channels, as well as by the use of genetic approaches to study protein function in vivo. More important, connexins have gained visibility because mutations in some connexin genes have been found to be linked to human genetic disorders. Connexin Methods and Protocols presents in detail a collection of te- niques currently used to study the cellular and molecular biology of connexins and their physiological properties. The field of gap junctions and connexin research has always been characterized by a multidisciplinary approach c- bining morphology, biochemistry, biophysics, and cellular and molecular biology. This book provides a series of cutting-edge protocols and includes a large spectrum of practical methods that are available to investigate the fu- tion of connexin channels. Connexin Methods and Protocols is divided into three main parts.

Complement Methods and Protocols
Complement Methods and Protocols

Author: B. Paul Morgan

Publisher: Springer Science & Business Media

Published: 2008-02-05

Total Pages: 268

ISBN-13: 159259056X

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The complement system, first described more than a century ago, was for many years the ugly duckling of the immunology world, but no more. Complement in recent years has blossomed into a fascinating and fast moving field of immediate relevance to clinical scientists in fields as diverse as transplantation biology, virology, and inflammation. Despite its emergence from the shadows, complement retains an unwarranted reputation for being “difficult.” This impression derives in large part from the superficially complicated nomenclature, a relic of the long and tortuous process of unraveling the system, of naming components in order of discovery rather than in a syst- atic manner. Once the barrier of nomenclature has been surmounted, then the true simplicity of the system becomes apparent. Complement comprises an activation system and a cytolytic system. The former has diverged to focus on complement to distinct targets—bacteria, - mune complexes, and others—so that texts now describe three activation pa- ways, closely related to one another, but each with some unique features. The cytolytic pathway is the same regardless of the activation process and kills cells by creating pores in the membrane. Complement plays an important role in killing bacteria and is essential for the proper handling of immune complexes. Problems occur when complement is activated in an inappropriate manner—the potent inflammation-inducing products of the cascade then cause unwanted tissue damage and destruction.

Eicosanoid Protocols
Eicosanoid Protocols

Author: Elias A Lianos

Publisher: Springer Science & Business Media

Published: 2008-02-03

Total Pages: 298

ISBN-13: 1592592635

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In Eicosanoid Protocols, Elias A. Lianos and a panel of hands-on experts present cutting-edge methods for the study of eicosanoids, including prostaglandins, thromboxanes, and leukotrienes. The readily reproducible methods described hereconcentrate on studying the regulation of expression and function of enzymes, particularly cyclooxygenase (and its two isoforms), phospholipase A2, and lipoxygenases involved in the synthesis of established eicosanoids. Additional chapters are devoted to the characterization and distribution of the thromboxane A2 receptor in tissues and the biological roles of novel eicosanoids. Timely and authoritative, the methods in this book will help their users in exploring the pathobiology of inflammation. Eicosanoid Protocols offers new and established researchers powerful, state-of-the-art tools to probe the regulation and function of eicosanoids.

Immunotoxin Methods and Protocols
Immunotoxin Methods and Protocols

Author: Walter A. Hall

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 305

ISBN-13: 1592591140

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Immunotoxins represent a new class of human therapeutics that have widespread applications and a potential that has not yet been fully recognized since they were first conceived of by Paul Ehrlich in 1906. The majority of advances in the development and implementation of immunotoxins has occurred over the last 20 years. The reasons for this use of immunotoxins in basic science and clinical research are the powerful concurrent advances in genetic engineering and receptor physiology. Recombinant technology has allowed investigators to produce sufficient quantities of a homogeneous c- pound that allows clinical trials to be performed. The identification of specific receptors on malignant cell types has enabled scientists to generate immunotoxins that have had positive results in clinical trials. As more cellular targets are identified in coming years, additional trials will be conducted in different disease states affecting still larger patient populations. Modulation of the immune system to decrease the humoral response to immunotoxins may improve their overall efficacy. As increasingly more effective compounds are generated, it will be necessary to decrease the local and systemic toxicity - sociated with these agents, and methods for doing so are presently being - veloped. The work presented in Immunotoxin Methods and Protocols focuses on three specific areas of immunotoxin investigation that are being conducted by experts throughout the world. The first section describes the construction and development of a variety of immunotoxins.