Peptide Research Protocols: Endothelin

ISBN-13: 9781617372919 / Angielski / Miękka / 2010 / 242 str.

Janet J. Maguire; Anthony P. Davenport

Opis

The endothelins are a remarkable family of signaling peptides: molecular biology predicted the existence of their receptors and synthetic enzymes prior to both the identification of the encoded proteins and the synthesis of antagonists and inhibitors for use as pharmacological tools. Although considerable advances have been made, culminating in the design of endothelin antagonists with the- peutic potential in cardiovascular disease, much remains to be discovered. Tantalizingly, new research frontiers are emerging. To support further progress, Peptide Research Protocols: Endothelin encompasses experimental protocols that interrogate all facets of an endogenous mammalian peptide s- tem, from peptide and receptor expression through synthetic pathway to peptide function and potential role in human disease. Chapters describe the use of molecular techniques to quantify the expression of mRNA for both endothelin receptors and the endothelin-converting enzymes. Peptides, precursors, receptors, and synthetic enzymes may be localized and quantified in plasma, culture supernatants, tissue homogenates, and tissue s- tions using antibodies, while additional information on receptor characterization may be obtained using radioligand binding techniques. Several protocols cover in vitro assays that determine the function of the endothelin peptides in isolated preparations, that characterize new endothelin receptor ligands, or provide inf- mation on the tissue-specific processing of endothelin precursor peptides.

Recenzje

"This book is excellent for what it intends to be: to provide a coherent series of protocols on a focused topic. It efficiently integrates biochemical, pharmacological, physiological and molecular biological methods..." -Acta Cardiologica

Spis treści

Part I. Endothelin Peptide Protocols Immunocytochemical Localization of Endothelin Peptides, Precursors, and Endothelin-Converting Enzymes Rhoda E. Kuc Analysis of Endothelins by Enzyme-Linked Immunosorbent Assay and Radioimmunoassay Anthony P. Davenport and Rhoda E. Kuc Separation of Endothelin Isoforms by High Performance Liquid Chromatography and Detection by Radioimmunoassay Michael J. Ashby Part II. Endothelin Receptor Protocols Radioligand Binding Assays and Quantitative Autoradiography of Endothelin Receptors Anthony P. Davenport and Rhoda E. Kuc Using Receptor Antagonists in Binding Studies to Characterize a Mammalian Endothelin Receptor J. Ruth Wu-Wong Quantification of Endothelin Receptor mRNA by Competitive RT-PCR Paula J. W. Smith and Juan Carlos Monge Part III. Endothelin-Converting Enzyme Protocols Detection and Assessment of Endothelin-Converting Enzyme Activity Carolyn D. Jackson and Anthony J. Turner Quantitative Measurement of mRNA Levels by RT-PCR: Studies of ECE-1 Isoforms Delphine M. Lees, Noorafza Q. Khan, Stewart Barker, and Roger Corder Evaluation of Endothelin-Converting Enzyme Inhibitors Using Cultured Cells, Roger Corder Endothelin-Converting Enzyme Activity in Vascular Smooth Muscle Preparations In Vitro Janet J. Maguire Part IV. In Vitro Functional Assays Characterization of a New Endothelin Receptor Ligand by In Vitro Assays J. Ruth Wu-Wong Use of In Vitro Organ Cultures of Human Saphenous Vein as a Model for Intimal Proliferation Karen E. Porter Part V. In Vivo Functional Assays Investigation of the Endothelin System in Experimental Heart Failure Gillian A. Gray and Lorcan Sherry Local Forearm Vasoconstriction to Endothelin-1 Measured by Venous Occlusion Plethysmography In Vivo Fiona E. Strachan and David J. Webb Index

Blurb

Tantalizing new research frontiers are opening in peptide research, where the endothelins constitute an important class of signals that can be manipulated therapeutically to treat cardiovascular disease. In Peptide Research Protocols: Endothelin, a panel of multidisciplinary experts describes in detail readily reproducible methods to investigate all aspects of peptide research by focusing on the endothelin system-from its synthesis and metabolism, to its function in health and disease. These methods use state-of-the-art molecular techniques to quantify the expression of mRNA, here using both endothelin receptors and the endothelin-converting enzymes. They show how peptides, precursors, receptors, and synthetic enzymes can be localized and quantified in plasma, culture supernatants, tissue homogenate, and tissue sections using antibodies, and how radioligand binding techniques can provide additional information on receptor characterization. In vitro assays are presented to determine the function of peptides in isolated preparations, to characterize new receptor ligands, and to provide information on the tissue-specific processing of precursor peptides-all in the endothelin system. Several in vivo protocols illustrate the role of the endothelin peptides in healthy human individuals and describe animal models that can be used to predict the therapeutic potential of cardiovascular drugs that manipulate endothelin synthesis or function.
Authoritative and timely, Peptide Research Protocols: Endothelin provides both a comprehensive introduction to endothelin biology and an up-to-date collection of proven techniques and experimental strategies for use in peptide research today.