title: A novel interaction of cGMP-dependent protein kinase I with troponin T.
abstract: cGMP-dependent protein kinase (cGK) is a major intracellular receptor of cGMP and is implicated in several signal transduction pathways. To identify proteins that participate in the cGMP/cGK signaling pathway, we employed the yeast two-hybrid system with cGK Ialpha as bait. cDNAs encoding slow skeletal troponin T (skTnT) were isolated from both mouse embryo and human skeletal muscle cDNA libraries. The skTnT protein interacted with cGK Ibeta but not with cGK II nor cAMP-dependent protein kinase. The yeast two-hybrid and in vitro binding assays revealed that the N-terminal region of cGK Ialpha, containing the leucine zipper motif, is sufficient for the association with skTnT. In vivo analysis, mutations in cGK Ialpha, which disrupted the leucine zipper motif, were shown to completely abolish the binding to skTnT. Furthermore, cGK I also interacted with cardiac TnT (cTnT) but not with cardiac troponin I (cTnI). Together with the observations that cTnI is a good substrate for cGK I and is effectively phosphorylated in the presence of cTnT in vitro, these findings suggest that TnT functions as an anchoring protein for cGK I and that cGK I may participate in the regulation of muscle contraction through phosphorylation of TnI.
authors.affiliation: Discovery Research Laboratory, Tanabe Seiyaku Co. Ltd., 2-50, Kawagishi 2-chome, Toda, Saitama 335-8505, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Yuasa
authors.forename: H
authors.initials: H
authors.lastname: Michibata
authors.forename: K
authors.initials: K
authors.lastname: Omori
authors.forename: N
authors.initials: N
authors.lastname: Yanaka
mesh_terms: D000818:Animals; D019556:COS Cells; D017869:Cyclic GMP-Dependent Protein Kinases / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D016350:Leucine Zippers; D051379:Mice; D009119:Muscle Contraction; D010766:Phosphorylation; D020107:Troponin T / Q000737:chemistry* / Q000378:metabolism
chemical_list: D020107:Troponin T; D017869:Cyclic GMP-Dependent Protein Kinases