Rho-kinase phosphorylates eNOS at threonine 495 in endothelial cells

Masayuki Sugimoto, Masanori Nakayama, Takaaki M. Goto, Mutsuki Amano, Kimihiro Komori, Kozo Kaibuchi

Research output: Contribution to journalArticlepeer-review

79 Citations (Scopus)


Endothelial nitric oxide synthase (eNOS) produces nitric oxide (NO), which is involved in various physiological functions of the cardiovascular system. eNOS is activated by dephosphorylation at Thr495 and phosphorylation at Ser1177. Inhibition of Rho-kinase, an effector of the small GTPase RhoA, leads to activation of Akt/PKB, which phosphorylates eNOS at Ser1177 and thereby promotes NO production. However, little is known about the effects of Rho-kinase on phosphorylation of Thr495. We here found that the constitutively active form of Rho-kinase phosphorylated eNOS at Thr495 in vitro. Expression of the constitutively active form of RhoA or Rho-kinase increased this phosphorylation in COS-7 cells. Addition of thrombin to cultured human umbilical vein endothelial cells induced phosphorylation of eNOS at Thr495. Treatment with Y27632, a Rho-kinase inhibitor, suppressed thrombin-induced phosphorylation at Thr495. These results indicate that Rho-kinase can directly phosphorylate eNOS at Thr495 to suppress NO production in endothelium.

Original languageEnglish
Pages (from-to)462-467
Number of pages6
JournalBiochemical and Biophysical Research Communications
Issue number2
Publication statusPublished - 21-09-2007
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology


Dive into the research topics of 'Rho-kinase phosphorylates eNOS at threonine 495 in endothelial cells'. Together they form a unique fingerprint.

Cite this