title: Methods for identification of cGKI substrates.
abstract: The cGMP-dependent protein kinases (cGK), which belong to the family of serine/threonine kinases, exhibit their diverse functions in cells through interaction with a variety of substrate proteins. Several substrates were identified and the interactions studied using different methods inter alia co-immunoprecipitation (Co-IP) and cGMP-agarose affinity purification. In the following chapter, we will describe the preparation of cell or tissue lysates, the procedures of cGMP-agarose affinity purification and co-immunoprecipitation, and finally the separation and analysis of the protein complexes by SDS-PAGE or mass spectrometry.
authors.affiliation: Pharmakologie und Toxikologie, Institut für Pharmazie, Universität Regensburg, Regensburg, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Salb
authors.forename: Jens
authors.initials: J
authors.lastname: Schlossmann
mesh_terms: D000818:Animals; D006152:Cyclic GMP / Q000737:chemistry / Q000378:metabolism; D062965:Cyclic GMP-Dependent Protein Kinase Type I / Q000737:chemistry* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D047468:Immunoprecipitation / Q000379:methods; D011485:Protein Binding; D013379:Substrate Specificity
chemical_list: D062965:Cyclic GMP-Dependent Protein Kinase Type I; D006152:Cyclic GMP