Hepatocyte growth factor activates endothelial nitric oxide synthase by Ca(2+)- and phosphoinositide 3-kinase/Akt-dependent phosphorylation in aortic endothelial cells

Kennedy Makondo; Kazuhiro Kimura; Naoki Kitamura; Takanori Kitamura; Daisuke Yamaji; Bae Dong Jung; Masayuki Saito

doi:10.1042/BJ20030326

Hepatocyte growth factor activates endothelial nitric oxide synthase by Ca(2+)- and phosphoinositide 3-kinase/Akt-dependent phosphorylation in aortic endothelial cells

Biochem J. 2003 Aug 15;374(Pt 1):63-9. doi: 10.1042/BJ20030326.

Authors

Kennedy Makondo¹, Kazuhiro Kimura, Naoki Kitamura, Takanori Kitamura, Daisuke Yamaji, Bae Dong Jung, Masayuki Saito

Affiliation

¹ Laboratory of Biochemistry, Department of Biomedical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan.

Abstract

Hepatocyte growth factor (HGF) causes endothelium-dependent vasodilation, but its relation to endothelial nitric oxide synthase (eNOS) activity remains to be elucidated. Treatment of bovine aortic endothelial cells with HGF increased eNOS activity within minutes, accompanied by an increase of activity-related site-specific phosphorylation of eNOS. The phosphorylation was completely abolished by pretreatment of the cells with a phosphoinositide 3-kinase (PI3K) inhibitor (wortmannin) and by transfection of dominant-negative Akt, and the enzyme activity was inhibited by wortmannin. In addition, eNOS activity and phosphorylation were abolished by pretreatment of the cells with an intracellular Ca(2+)-chelator, bis-(o-aminophenoxy)ethane-N,N,N',N'-tetra-acetic acid tetrakis(acetoxymethyl ester) (BAPTA/AM), with a suppression of Akt phosphorylation. These results suggest that HGF stimulates eNOS activity by a PI3K/Akt-dependent phosphorylation in a Ca(2+)-sensitive manner in vascular endothelial cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Analysis of Variance
Animals
Aorta
Calcium / metabolism*
Cattle
Cells, Cultured
Endothelium, Vascular / enzymology*
Enzyme Activation / drug effects
Hepatocyte Growth Factor / pharmacology*
Humans
Kinetics
Mitogen-Activated Protein Kinase 1 / metabolism
Mitogen-Activated Protein Kinase 3
Mitogen-Activated Protein Kinases / metabolism
Nitric Oxide Synthase / metabolism*
Nitric Oxide Synthase Type III
Phosphatidylinositol 3-Kinases / metabolism*
Protein Serine-Threonine Kinases*
Protein-Tyrosine Kinases / metabolism
Proto-Oncogene Proteins / metabolism*
Proto-Oncogene Proteins c-akt
Recombinant Proteins / pharmacology

Substances

Proto-Oncogene Proteins
Recombinant Proteins
Hepatocyte Growth Factor
NOS3 protein, human
Nitric Oxide Synthase
Nitric Oxide Synthase Type III
Protein-Tyrosine Kinases
AKT1 protein, human
Protein Serine-Threonine Kinases
Proto-Oncogene Proteins c-akt
Mitogen-Activated Protein Kinase 1
Mitogen-Activated Protein Kinase 3
Mitogen-Activated Protein Kinases
Calcium