Inactivation of Ca2+/calmodulin-dependent protein kinase I by S-glutathionylation of the active-site cysteine residue.
We show that Ca(2+)/calmodulin(CaM)-dependent protein kinase I (CaMKI) is directly inhibited by its S-glutathionylation at the Cys(179). In vitro studies demonstrated that treatment of CaMKI with diamide and glutathione results in inactivation of the enzyme, with a concomitant S-glutathionylation of CaMKI at Cys(179) detected by mass spectrometry. Mutagenesis studies confirmed that S-glutathionylation of Cys(179) is both necessary and sufficient for the inhibition of CaMKI by diamide and glutathione. In transfected cells expressing CaMKI, treatment with diamide caused a reversible decrease in CaMKI activity. Cells expressing mutant CaMKI (179CV) proved resistant in this regard. Thus, our results indicate that the reversible regulation of CaMKI via its modification at Cys(179) is an important mechanism in processing calcium signal transduction in cells.
Kambe
Toshie
T
Department of Pharmacology, High Technology Research Center, Showa Pharmaceutical University, Machida, Tokyo, Japan.
Song
Tao
T
Takata
Tsuyoshi
T
Hatano
Naoya
N
Miyamoto
Yoshiaki
Y
Nozaki
Naohito
N
Naito
Yasuhito
Y
Tokumitsu
Hiroshi
H
Watanabe
Yasuo
Y
D001665:Binding Sites / Q000235:genetics; D002118:Calcium / Q000378:metabolism*; D054729:Calcium-Calmodulin-Dependent Protein Kinase Type 1 / Q000737:chemistry / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism*; D003545:Cysteine / Q000737:chemistry*; D003958:Diamide / Q000494:pharmacology; D005978:Glutathione / Q000378:metabolism*; D007477:Ions / Q000378:metabolism; D013058:Mass Spectrometry; D011494:Protein Kinases / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects
D002147:Calmodulin; D007477:Ions; D003958:Diamide; D011494:Protein Kinases; D054729:Calcium-Calmodulin-Dependent Protein Kinase Type 1; D005978:Glutathione; D003545:Cysteine; D002118:Calcium