Domain Mapping Studies Reveal That the M Domain of hsp90 Serves as a Molecular Scaffold to Regulate Akt-Dependent Phosphorylation of Endothelial Nitric Oxide Synthase and NO Release

DOI Web Site 被引用文献11件
  • Jason Fontana
    From the Department of Pharmacology and Molecular Cardiobiology Program (J.F., D.F., Y.C., T.A.F., T.J.M., W.C.S.), Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, Conn; and the Institute of Molecular and Cellular Biosciences (N.F., T.T.), Bunkyo-ku, Tokyo, Japan.
  • David Fulton
    From the Department of Pharmacology and Molecular Cardiobiology Program (J.F., D.F., Y.C., T.A.F., T.J.M., W.C.S.), Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, Conn; and the Institute of Molecular and Cellular Biosciences (N.F., T.T.), Bunkyo-ku, Tokyo, Japan.
  • Yan Chen
    From the Department of Pharmacology and Molecular Cardiobiology Program (J.F., D.F., Y.C., T.A.F., T.J.M., W.C.S.), Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, Conn; and the Institute of Molecular and Cellular Biosciences (N.F., T.T.), Bunkyo-ku, Tokyo, Japan.
  • Todd A. Fairchild
    From the Department of Pharmacology and Molecular Cardiobiology Program (J.F., D.F., Y.C., T.A.F., T.J.M., W.C.S.), Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, Conn; and the Institute of Molecular and Cellular Biosciences (N.F., T.T.), Bunkyo-ku, Tokyo, Japan.
  • Timothy J. McCabe
    From the Department of Pharmacology and Molecular Cardiobiology Program (J.F., D.F., Y.C., T.A.F., T.J.M., W.C.S.), Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, Conn; and the Institute of Molecular and Cellular Biosciences (N.F., T.T.), Bunkyo-ku, Tokyo, Japan.
  • Naoya Fujita
    From the Department of Pharmacology and Molecular Cardiobiology Program (J.F., D.F., Y.C., T.A.F., T.J.M., W.C.S.), Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, Conn; and the Institute of Molecular and Cellular Biosciences (N.F., T.T.), Bunkyo-ku, Tokyo, Japan.
  • Takashi Tsuruo
    From the Department of Pharmacology and Molecular Cardiobiology Program (J.F., D.F., Y.C., T.A.F., T.J.M., W.C.S.), Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, Conn; and the Institute of Molecular and Cellular Biosciences (N.F., T.T.), Bunkyo-ku, Tokyo, Japan.
  • William C. Sessa
    From the Department of Pharmacology and Molecular Cardiobiology Program (J.F., D.F., Y.C., T.A.F., T.J.M., W.C.S.), Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, Conn; and the Institute of Molecular and Cellular Biosciences (N.F., T.T.), Bunkyo-ku, Tokyo, Japan.

抄録

<jats:p>Protein-protein interactions with the molecular chaperone hsp90 and phosphorylation on serine 1179 by the protein kinase Akt leads to activation of endothelial nitric oxide synthase. However, the interplay between these protein-protein interactions remains to be established. In the present study, we show that vascular endothelial growth factor stimulates the coordinated association of hsp90, Akt, and resultant phosphorylation of eNOS. Characterization of the domains of hsp90 required to bind eNOS, using yeast 2-hybrid, cell-based coprecipitation experiments, and GST-fusion proteins, revealed that the M region of hsp90 interacts with the amino terminus of eNOS and Akt. The addition of purified hsp90 to in vitro kinase assays facilitates Akt-driven phosphorylation of recombinant eNOS protein, but not a short peptide encoding the Akt phosphorylation site, suggesting that hsp90 may function as a scaffold for eNOS and Akt. In vivo, coexpression of adenoviral or the cDNA for hsp90 with eNOS promotes nitric oxide release; an effect eliminated using a catalytically functional phosphorylation mutant of eNOS. These results demonstrate that stimulation of endothelial cells with vascular endothelial growth factor recruits eNOS and Akt to an adjacent region on the same domain of hsp90, thereby facilitating eNOS phosphorylation and enzyme activation.</jats:p>

収録刊行物

  • Circulation Research

    Circulation Research 90 (8), 866-873, 2002-05-03

    Ovid Technologies (Wolters Kluwer Health)

被引用文献 (11)*注記

もっと見る

キーワード

詳細情報 詳細情報について

問題の指摘

ページトップへ