Calcium/calmodulin-dependent protein kinase I inhibits neuronal nitric-oxide synthase activity through serine 741 phosphorylation

Tao Song, Naoya Hatano, Mariko Horii, Hiroshi Tokumitsu, Fuminori Yamaguchi, Masaaki Tokuda, Yasuo Watanabe

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

We demonstrate here that neuronal nitric-oxide synthase (nNOS) is phosphorylated and inhibited by a constitutively active form of Ca 2+/calmodulin (CaM)-dependent protein kinase I (CaM-K I1-293). Substitution of Ser741 to Ala in nNOS blocked the phosphorylation and the inhibitory effect. Mimicking phosphorylation at Ser741 by Ser to Asp mutation resulted in decreased binding of and activation by CaM, since the mutation was within the CaM-binding domain. CaM-K I1-293 gave phosphorylation of nNOS at Ser741 in transfected cells, resulting in 60-70% inhibition of nNOS activity. Wild-type CaM-K I also did phosphorylate nNOS at Ser 741 in transfected cells, but either CaM-K II or CaM-K IV did not. These results raise the possibility of a novel cross-talk between nNOS and CaM-K I through the phosphorylation of Ser741 on nNOS.

Original languageEnglish
Pages (from-to)133-137
Number of pages5
JournalFEBS Letters
Volume570
Issue number1-3
DOIs
Publication statusPublished - Jul 16 2004
Externally publishedYes

Fingerprint

Calcium-Calmodulin-Dependent Protein Kinase Type 1
Nitric Oxide Synthase Type I
Calcium-Calmodulin-Dependent Protein Kinases
Phosphorylation
Calmodulin
Serine
Mutation
Substitution reactions
Chemical activation

Keywords

  • CaM, calmodulin
  • CaM-K I, II, and IV, calcium/calmodulin-dependent protein kinases I, II, and IV
  • CaM-KK, calcium/calmodulin-dependent protein kinase kinase
  • nNOS, neuronal nitric-oxide synthase
  • PP2A, protein phosphatase 2A

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Calcium/calmodulin-dependent protein kinase I inhibits neuronal nitric-oxide synthase activity through serine 741 phosphorylation. / Song, Tao; Hatano, Naoya; Horii, Mariko; Tokumitsu, Hiroshi; Yamaguchi, Fuminori; Tokuda, Masaaki; Watanabe, Yasuo.

In: FEBS Letters, Vol. 570, No. 1-3, 16.07.2004, p. 133-137.

Research output: Contribution to journalArticle

Song, Tao ; Hatano, Naoya ; Horii, Mariko ; Tokumitsu, Hiroshi ; Yamaguchi, Fuminori ; Tokuda, Masaaki ; Watanabe, Yasuo. / Calcium/calmodulin-dependent protein kinase I inhibits neuronal nitric-oxide synthase activity through serine 741 phosphorylation. In: FEBS Letters. 2004 ; Vol. 570, No. 1-3. pp. 133-137.
@article{c413a619ed40429daa26339b53be4747,
title = "Calcium/calmodulin-dependent protein kinase I inhibits neuronal nitric-oxide synthase activity through serine 741 phosphorylation",
abstract = "We demonstrate here that neuronal nitric-oxide synthase (nNOS) is phosphorylated and inhibited by a constitutively active form of Ca 2+/calmodulin (CaM)-dependent protein kinase I (CaM-K I1-293). Substitution of Ser741 to Ala in nNOS blocked the phosphorylation and the inhibitory effect. Mimicking phosphorylation at Ser741 by Ser to Asp mutation resulted in decreased binding of and activation by CaM, since the mutation was within the CaM-binding domain. CaM-K I1-293 gave phosphorylation of nNOS at Ser741 in transfected cells, resulting in 60-70{\%} inhibition of nNOS activity. Wild-type CaM-K I also did phosphorylate nNOS at Ser 741 in transfected cells, but either CaM-K II or CaM-K IV did not. These results raise the possibility of a novel cross-talk between nNOS and CaM-K I through the phosphorylation of Ser741 on nNOS.",
keywords = "CaM, calmodulin, CaM-K I, II, and IV, calcium/calmodulin-dependent protein kinases I, II, and IV, CaM-KK, calcium/calmodulin-dependent protein kinase kinase, nNOS, neuronal nitric-oxide synthase, PP2A, protein phosphatase 2A",
author = "Tao Song and Naoya Hatano and Mariko Horii and Hiroshi Tokumitsu and Fuminori Yamaguchi and Masaaki Tokuda and Yasuo Watanabe",
year = "2004",
month = "7",
day = "16",
doi = "10.1016/j.febslet.2004.05.083",
language = "English",
volume = "570",
pages = "133--137",
journal = "FEBS Letters",
issn = "0014-5793",
publisher = "Elsevier",
number = "1-3",

}

TY - JOUR

T1 - Calcium/calmodulin-dependent protein kinase I inhibits neuronal nitric-oxide synthase activity through serine 741 phosphorylation

AU - Song, Tao

AU - Hatano, Naoya

AU - Horii, Mariko

AU - Tokumitsu, Hiroshi

AU - Yamaguchi, Fuminori

AU - Tokuda, Masaaki

AU - Watanabe, Yasuo

PY - 2004/7/16

Y1 - 2004/7/16

N2 - We demonstrate here that neuronal nitric-oxide synthase (nNOS) is phosphorylated and inhibited by a constitutively active form of Ca 2+/calmodulin (CaM)-dependent protein kinase I (CaM-K I1-293). Substitution of Ser741 to Ala in nNOS blocked the phosphorylation and the inhibitory effect. Mimicking phosphorylation at Ser741 by Ser to Asp mutation resulted in decreased binding of and activation by CaM, since the mutation was within the CaM-binding domain. CaM-K I1-293 gave phosphorylation of nNOS at Ser741 in transfected cells, resulting in 60-70% inhibition of nNOS activity. Wild-type CaM-K I also did phosphorylate nNOS at Ser 741 in transfected cells, but either CaM-K II or CaM-K IV did not. These results raise the possibility of a novel cross-talk between nNOS and CaM-K I through the phosphorylation of Ser741 on nNOS.

AB - We demonstrate here that neuronal nitric-oxide synthase (nNOS) is phosphorylated and inhibited by a constitutively active form of Ca 2+/calmodulin (CaM)-dependent protein kinase I (CaM-K I1-293). Substitution of Ser741 to Ala in nNOS blocked the phosphorylation and the inhibitory effect. Mimicking phosphorylation at Ser741 by Ser to Asp mutation resulted in decreased binding of and activation by CaM, since the mutation was within the CaM-binding domain. CaM-K I1-293 gave phosphorylation of nNOS at Ser741 in transfected cells, resulting in 60-70% inhibition of nNOS activity. Wild-type CaM-K I also did phosphorylate nNOS at Ser 741 in transfected cells, but either CaM-K II or CaM-K IV did not. These results raise the possibility of a novel cross-talk between nNOS and CaM-K I through the phosphorylation of Ser741 on nNOS.

KW - CaM, calmodulin

KW - CaM-K I, II, and IV, calcium/calmodulin-dependent protein kinases I, II, and IV

KW - CaM-KK, calcium/calmodulin-dependent protein kinase kinase

KW - nNOS, neuronal nitric-oxide synthase

KW - PP2A, protein phosphatase 2A

UR - http://www.scopus.com/inward/record.url?scp=3142528210&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=3142528210&partnerID=8YFLogxK

U2 - 10.1016/j.febslet.2004.05.083

DO - 10.1016/j.febslet.2004.05.083

M3 - Article

C2 - 15251453

AN - SCOPUS:3142528210

VL - 570

SP - 133

EP - 137

JO - FEBS Letters

JF - FEBS Letters

SN - 0014-5793

IS - 1-3

ER -