Peptide Research Protocols: Endothelin

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Juni 2002



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.


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



"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
EAN: 9780896039933
ISBN: 0896039935
Untertitel: Endothelin. 'Methods in Molecular Biology'. 2002. Auflage. Sprache: Englisch.
Erscheinungsdatum: Juni 2002
Seitenanzahl: 242 Seiten
Format: gebunden
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