title: Identification of a conserved residue responsible for the autoinhibition of cGMP-dependent protein kinase Ialpha and beta.
abstract: We isolated a constitutively active form of cGMP-dependent protein kinase Ialpha (cGK Ialpha) by PCR-driven random mutagenesis. The replacement of Ile-63 by Thr in the autoinhibitory domain results in the enhancement of autophosphorylation and the basal kinase activity in the absence of cGMP. The hydrophobicity at position 63 is essential for the inactive state of cGK Ialpha, and Ile-78 of cGK Ibeta is also required for the autoinhibitory property. Furthermore, cGK Ialpha (Ile-63-Thr) is constitutively active in vivo. These findings suggest that a conserved residue in the autoinhibitory domain was involved in the autoinhibition of both cGK Is.
authors.affiliation: Discovery Research Laboratory, Tanabe Seiyaku Co. Ltd., 2-50, Kawagishi 2-chome, Toda, Saitama, 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: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D020134:Catalytic Domain / Q000235:genetics; D002417:Cattle; D017124:Conserved Sequence; D062965:Cyclic GMP-Dependent Protein Kinase Type I; D017869:Cyclic GMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors* / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D006801:Humans; D066298:In Vitro Techniques; D016296:Mutagenesis; D010766:Phosphorylation; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism
chemical_list: D017931:DNA Primers; D011994:Recombinant Proteins; D062965:Cyclic GMP-Dependent Protein Kinase Type I; D017869:Cyclic GMP-Dependent Protein Kinases; C569718:PRKG1 protein, human