Title Phosphodiesterase methods and protocols / edited by Claire Lugnier

Published Totowa, N.J. : Humana Press, ©2005

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Description 1 online resource (xii, 324 pages) : illustrations Series Methods in molecular biology, 1064-3745 ; v. 307 Methods in molecular biology (Clifton, N.J.) ; v. 307 Contents Study of cAMP microdomains in cells / Marco Mongillo [and others] -- High-resolution measurements of cAMP signals in three-dimensional microdomains / Jeffrey W. Karpen and Thomas C. Rich -- Cygnets: in vivo characterization of novel cGMP indicators and in vivo imaging of intracellular cGMP / Akira Honda [and others] -- High-throughput screening of PDE activity in living cells / Thomas C. Rich and Jeffrey W. Karpen -- Assessment of PDE isozyme contribution in cell and tissue extracts / Thäråse Keravis, Rima Thaseldar-Roumiä, and Claire Lugnier -- Localization of the cGMP-hydrolysing phosphodiesterase type 9 in the rat brain by a non-radioactive in situ hybridization / Wilma C.G. Van Staveren and Marjanne Markerink-Van Ittersum -- Determination of Ca2+/calmodulin-stimulated phosphodiesterase activity in intact cells / C. Yan -- Adenovirus mediated overexpression of murine cyclic nucleotide phosphodiesterase 3B (mPDE3B) / Faiyaz Ahmad [and others] -- Identification of promoter elements in the 5'-flanking region of the murine cyclic nucleotide phosphodiesterase 3B gene / Hanguan Liu [and others] Purification of PDE6 isozymes from mammalian retina / Dana C. Pentia [and others] -- cGMP binding to the regulatory GAF domains of photoreceptor phosphodiesterase / Rick H. Cote -- Renaturation of the catalytic domain of PDE4A expressed in escherichia coli as inclusion bodies / Wito Richter, Thomas Hermsdorf, and Dietrich Dettmer -- Determining the subunit structure of phosphodiesterases using gel-filtration and sucrose density gradient centrifugation / Wito Richter -- Crystallization of cyclic nucleotide phosphodiesterases / Hengming Ke, Qing Huai, and Robert X. Xu -- Generation of PDE4 knockout mice by gene targeting / S.L. Catherine Jin, Anne M. Latour, and Marco Conti -- Immunoprecipitating PDE2 phosphorylated and inactivated by associated protein kinase / J. Kelley Bentley -- Methods to investigate ERK MAP kinase phosphorylation and regulation of activity of PDE4 cyclicamp-specific phosphodiesterases / Elaine V. Hill, Miles D. Houslay, and George S. Baillie -- R Radiolabeled ligand binding to the catalytic sites or allosteric sites of PDE5 and PDE11 / James L. Weeks [and others] -- Analysis of dimerization determinants of the PDE6 catalytic subunits / Khakim G. Muradov, Kimberley K. Boyd, and Nikolai O. Artemyev -- Interaction between catalytic and inhibitory subunits of PDE6 / Nikolai O. Artemyev -- Purification, reconstitution on lipid vesciles and assays of PDE6 and its activator G protein, transducin -- Theodore G. Wensel, Feng He, and Justine A. Malinski Summary The multigenic family of cyclic nucleotide phosphodiesterases (PDEs) is now known to offer many new therapeutic possibilities through their influence over intracellular signaling, though their precise cell mechanisms to modulating that process remain uncertain. In Phosphodiesterase Methods and Protocols, a panel of research leaders in the PDE field describes new concepts and techniques for investigating the role of PDEs in orchestrating normal and pathophysiological responses. Presented in step-by-step detail, these readily reproducible methods allow the measurement of cyclic nucleotide variations in living cells, as well as their visualization in a spatiotemporal manner, the localization and characterization of their activities in tissues and living cells, and the assessment of targeted PDEs in creating specific tools and drugs. Additional chapters discuss the generation of PDE4 knockout mice to demonstrate not only the potential role of targeted PDEs, but also their use in further studies of the central role of PDE regulation in intracellular signaling control. These techniques offer clinicians a way to find new therapies for numerous pathologies whose molecular origins are unknown and whose treatment is symptomatic. Alterations of intracellular signaling related to PDE deregulation in such pathologies as inflammation, neurodegeneration, and cancer may explain the difficulties observed in their prevention and treatment. The protocols follow the successful Methods in Molecular Biology series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Path-breaking and highly practical, Phosphodiesterase Methods and Protocols offers cell biologists, molecular biologists, pharmacologists, and medicinal chemists a broad ranging survey of the advanced tools and concepts needed to understand the role of PDEs in physiological functions, their implications in several critical pathologies, and the opportunities they offer for new drug design Bibliography Includes bibliographical references and index Notes English Print version record Subject Phosphodiesterases -- Laboratory manuals Esterases. Genetic engineering. Life sciences. Physical sciences. Genetics -- Technique. Hydrolases. Enzymes. Methodology. Physiology. Protein engineering. Biological assay. Phosphoric Diester Hydrolases -- analysis Phosphoric Diester Hydrolases -- physiology Biological Assay -- methods Protein Engineering -- methods Publication Formats Esterases Investigative Techniques Genetic Engineering Biological Science Disciplines Natural Science Disciplines Genetic Techniques Analytical, Diagnostic and Therapeutic Techniques and Equipment Hydrolases Publication Characteristics Enzymes Disciplines and Occupations Enzymes and Coenzymes Chemicals and Drugs Phosphoric Diester Hydrolases Methods Physiology Protein Engineering Biological Assay biological sciences. physical sciences. enzyme. methodology. physiology. MEDICAL -- Biochemistry. Protein engineering Physiology Physical sciences Methodology Life sciences Hydrolases Genetics -- Technique Genetic engineering Esterases Enzymes Biological assay Phosphodiesterases Genre/Form Laboratory manuals Laboratory manuals. Manuels de laboratoire. Form Electronic book Author Lugnier, Claire ISBN 1588293149 9781588293145 9781592598397 1592598390 159259901X 9781592599011

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