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.
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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.
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