title: KN-62, a specific Ca++/calmodulin-dependent protein kinase inhibitor, reversibly depresses the rate of beating of cultured fetal mouse cardiac myocytes.
abstract: Effects of KN-62 (1-[N,O-bis(5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4- phenylpiperazine), a specific Ca++/calmodulin (CaM)-dependent protein kinase inhibitor, were examined on the rate of spontaneous beating and the intracellular Ca++ transient of cultured myocytes from fetal mouse ventricle. KN-62 depressed the rate of beating in a dose-dependent fashion. Spontaneous beating ceased 10 min after the administration of 1 microM KN-62 and recovered gradually after washing with cultured medium. Addition of KN-04 [N-(1-1[P-(5-isoquinolinsulfonyl)benzyl]-2-(4- phenylpiperazinyl)ethyl)-5-isoquinolinsulfonamide; 1 microM], an analog of KN-62, did not change the rate of beating. In the experiment using an intracellular Ca++ fluorescence indicator, fluo-3, KN-62 depressed the fluo-3 intensity at a systolic phase. The kinase activity to syntide-2 of Ca++/CaM kinase II purified from the rabbit heart was inhibited by KN-62, but not by KN-04. Addition of KN-62 inhibited the phosphorylation of phospholamban by Ca++/CaM kinase II in a dose-dependent manner. KN-62 depressed the Ca(++)-pumping ATPase activity in the presence of Ca++ and CaM by 32%. These findings indicate that Ca++/CaM kinase II changes an intracellular Ca++ transient and modulates the rate of beating at least in part.
authors.affiliation: Department of Pharmacology, Nagoya University School of Medicine, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Okazaki
authors.forename: T
authors.initials: T
authors.lastname: Ishikawa
authors.forename: M
authors.initials: M
authors.lastname: Inui
authors.forename: M
authors.initials: M
authors.lastname: Tada
authors.forename: K
authors.initials: K
authors.lastname: Goshima
authors.forename: T
authors.initials: T
authors.lastname: Okamoto
authors.forename: H
authors.initials: H
authors.lastname: Hidaka
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives*; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors*; D000252:Calcium-Transporting ATPases / Q000378:metabolism; D002478:Cells, Cultured; D006321:Heart / Q000187:drug effects* / Q000196:embryology / Q000502:physiology; D006339:Heart Rate / Q000187:drug effects*; D007546:Isoquinolines / Q000494:pharmacology*; D051379:Mice; D008813:Mice, Inbred ICR; D009206:Myocardium / Q000201:enzymology; D010766:Phosphorylation; D010879:Piperazines / Q000494:pharmacology*; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000201:enzymology; D000099211:Phospholamban
chemical_list: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D002118:Calcium; D002135:Calcium-Binding Proteins; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D000252:Calcium-Transporting ATPases; D007546:Isoquinolines; D010879:Piperazines; D000099211:Phospholamban; C063302:KN 62