The mechanisms of NO inhibition of CaMK [Ca2+/CaM (calmodulin)-dependent protein kinase] II activity were studied. In rat pituitary tumour GH3 cells, TRH [thyrotrophin (TSH)-releasing hormone]-stimulated phosphorylation of nNOS [neuronal NOS (NO synthase)] at Ser847 was sensitive to an inhibitor of CaMKs, KN-93, and was enhanced by inhibition of nNOS with 7NI (7-nitroindazole). Enzyme activity of CaMKII following in situ treatment with 7NI was also increased. The in vitro activity of CaMKII was inhibited by co-incubation either with nNOS and L-arginine or with NO donors SNAP (S-nitroso-N-acetyl-DL-penicillamine) and DEA-NONOate [diethylamine-NONOate (diazeniumdiolate)]. Once inhibited by these treatments, CaMKII was observed to undergo full reactivation on the addition of a reducing reagent, DTT (dithiothreitol). In transfected cells expressing CaMKII and nNOS, treatment with the calcium ionophore A23187 further revealed nNOS phosphorylation at Ser847, which was enhanced by 7NI and CaMKII S-nitrosylation. Mutated CaMKII (C6A), in which Cys6 was substituted with an alanine residue, was refractory to 7NI-induced enhancement of nNOS phosphorylation or to CaMKII S-nitrosylation. Furthermore, we could identify Cys6 as a direct target for S-nitrosylation of CaMKII using MS. In addition, treatment with glutamate caused an increase in CaMKII S-nitrosylation in rat hippocampal slices. This glutamate-induced S-nitrosylation was blocked by 7NI. These results suggest that inactivation of CaMKII mediated by S-nitrosylation at Cys6 may contribute to NO-induced neurotoxicity in the brain.
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Research Article|
May 14 2008
Nitric oxide-mediated modulation of calcium/calmodulin-dependent protein kinase II
Tao Song;
Tao Song
*Department of Pharmacology, Showa Pharmaceutical University, Machida, Tokyo 194-8543, Japan
†Department of Cell Physiology, Faculty of Medicine, Kagawa University, Kagawa 761-0793, Japan
‡Department of Anesthesiology, The First Affiliated Hospital, China Medical University, Shenyang 110001, China
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Naoya Hatano;
Naoya Hatano
†Department of Cell Physiology, Faculty of Medicine, Kagawa University, Kagawa 761-0793, Japan
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Toshie Kambe;
Toshie Kambe
*Department of Pharmacology, Showa Pharmaceutical University, Machida, Tokyo 194-8543, Japan
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Yoshiaki Miyamoto;
Yoshiaki Miyamoto
*Department of Pharmacology, Showa Pharmaceutical University, Machida, Tokyo 194-8543, Japan
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Hideshi Ihara;
Hideshi Ihara
§Department of Biological Science, Graduate School of Science, Osaka Prefecture University, Sakai, Osaka 599-8531, Japan
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Hideyuki Yamamoto;
Hideyuki Yamamoto
∥Department of Biochemistry, School of Medicine, University of the Ryukyus, Nishihara, Okinawa 903-0215, Japan
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Katsuyoshi Sugimoto;
Katsuyoshi Sugimoto
†Department of Cell Physiology, Faculty of Medicine, Kagawa University, Kagawa 761-0793, Japan
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Kodai Kume;
Kodai Kume
†Department of Cell Physiology, Faculty of Medicine, Kagawa University, Kagawa 761-0793, Japan
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Fuminori Yamaguchi;
Fuminori Yamaguchi
†Department of Cell Physiology, Faculty of Medicine, Kagawa University, Kagawa 761-0793, Japan
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Masaaki Tokuda;
Masaaki Tokuda
†Department of Cell Physiology, Faculty of Medicine, Kagawa University, Kagawa 761-0793, Japan
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Yasuo Watanabe
Yasuo Watanabe
1
*Department of Pharmacology, Showa Pharmaceutical University, Machida, Tokyo 194-8543, Japan
†Department of Cell Physiology, Faculty of Medicine, Kagawa University, Kagawa 761-0793, Japan
1To whom correspondence should be addressed at the present address: Department of Pharmacology, Showa Pharmaceutical University, Machida, Tokyo 194-8543, Japan (email yasuwata@ac.shoyaku.ac.jp).
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Publisher: Portland Press Ltd
Received:
August 31 2007
Revision Received:
February 13 2008
Accepted:
February 14 2008
Accepted Manuscript online:
February 14 2008
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2008 Biochemical Society
2008
Biochem J (2008) 412 (2): 223–231.
Article history
Received:
August 31 2007
Revision Received:
February 13 2008
Accepted:
February 14 2008
Accepted Manuscript online:
February 14 2008
Citation
Tao Song, Naoya Hatano, Toshie Kambe, Yoshiaki Miyamoto, Hideshi Ihara, Hideyuki Yamamoto, Katsuyoshi Sugimoto, Kodai Kume, Fuminori Yamaguchi, Masaaki Tokuda, Yasuo Watanabe; Nitric oxide-mediated modulation of calcium/calmodulin-dependent protein kinase II. Biochem J 1 June 2008; 412 (2): 223–231. doi: https://doi.org/10.1042/BJ20071195
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