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Brain ischemia induces serine phosphorylation of neuronal nitric oxide synthase by Ca(2+)/calmodulin-dependent protein kinase II in rat hippocampus.

AbstractAIM:
To investigate whether brain ischemia induces serine phosphorylation of neuronal nitric oxide synthase (nNOS) by Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) and the interaction between CaMKII? and nNOS in rat hippocampus.
METHODS:
Brain ischemia was induced by bilateral carotid artery occlusion procedure. Phosphorylation and the interaction of proteins were studied by immunoprecipitation and immunoblotting. We investigated during brain ischemia serine phosphorylation and amount of nNOS in crude membranes fraction (P) and cytosolic fraction (S), interaction between CaMKIIalpha and nNOS, and the effects of 1-[N,O-bis-(5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine (KN-62, a selective inhibitor of CaMKII) on phosphorylation and the interaction of proteins in P.
RESULTS:
Serine phosphorylation of nNOS in P increased persistently during brain ischemia, and 15 min ischemia-induced serine phosphorylation of nNOS was attenuated significantly by KN-62. But there was no serine phosphorylation of nNOS in S. The distributions of nNOS were not affected by ischemia and KN-62. However, the binding levels of both CaMKIIalpha with nNOS and Thr(286) autophosphorylated CaMKIIalpha with nNOS increased after ischemia, and were diminished by KN-62.
CONCLUSION:
CaMKII interacted with nNOS and regulated serine phosphorylation of nNOS during brain ischemia.
AuthorsXue-bo Yan, Fan-jie Meng, Bo Song, Guang-yi Zhang
JournalActa pharmacologica Sinica (Acta Pharmacol Sin) Vol. 25 Issue 5 Pg. 617-22 (May 2004) ISSN: 1671-4083 [Print] United States
PMID15132828 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Serine
  • KN 62
  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
  • Nitric Oxide Synthase
  • Nitric Oxide Synthase Type I
  • Nos1 protein, rat
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2
  • Calcium-Calmodulin-Dependent Protein Kinases
Topics
  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine (analogs & derivatives, pharmacology)
  • Animals
  • Brain Ischemia (enzymology, metabolism)
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2
  • Calcium-Calmodulin-Dependent Protein Kinases (antagonists & inhibitors, metabolism)
  • Hippocampus (enzymology)
  • Male
  • Nitric Oxide Synthase (metabolism)
  • Nitric Oxide Synthase Type I
  • Phosphorylation
  • Rats
  • Rats, Sprague-Dawley
  • Serine (metabolism)

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