Circulation Research. 2002;90:866-873
Published online before print March 28, 2002, doi: 10.1161/01.RES.0000016837.26733.BE
Published online before print March 28, 2002, doi: 10.1161/01.RES.0000016837.26733.BE
© 2002 American Heart Association, Inc.
Molecular Medicine |
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
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.
Correspondence to William C. Sessa, PhD, Dept of Pharmacology and Molecular Cardiobiology Program, Boyer Center for Molecular Medicine, Yale University School of Medicine, 295 Congress Ave, New Haven, CT 06536. E-mail william.sessa{at}yale.edu
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.
Key Words: nitric oxide • signaling • scaffold • hsp90 • Akt
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|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
 |
|
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|
|
|
|
 |
|
 A. Martinez-Ruiz, L. Villanueva, C. G. de Orduna, D. Lopez-Ferrer, M. A. Higueras, C. Tarin, I. Rodriguez-Crespo, J. Vazquez, and S. Lamas S-nitrosylation of Hsp90 promotes the inhibition of its ATPase and endothelial nitric oxide synthase regulatory activities PNAS, June 14, 2005; 102(24): 8525 - 8530. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Herrera and J. L. Garvin Recent Advances in the Regulation of Nitric Oxide in the Kidney Hypertension, June 1, 2005; 45(6): 1062 - 1067. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Q. Wei and Y. Xia Roles of 3-Phosphoinositide-dependent Kinase 1 in the Regulation of Endothelial Nitric-oxide Synthase Phosphorylation and Function by Heat Shock Protein 90 J. Biol. Chem., May 6, 2005; 280(18): 18081 - 18086. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
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|
|
|
|
 |
|
 T. J. Stalker, Y. Gong, and R. Scalia The Calcium-Dependent Protease Calpain Causes Endothelial Dysfunction in Type 2 Diabetes Diabetes, April 1, 2005; 54(4): 1132 - 1140. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Terasawa, M. Minami, and Y. Minami Constantly Updated Knowledge of Hsp90 J. Biochem., April 1, 2005; 137(4): 443 - 447. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Connelly, M. Madhani, and A. J. Hobbs Resistance to Endotoxic Shock in Endothelial Nitric-oxide Synthase (eNOS) Knock-out Mice: A PRO-INFLAMMATORY ROLE FOR eNOS-DERIVED NO IN VIVO J. Biol. Chem., March 18, 2005; 280(11): 10040 - 10046. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Bucci, F. Roviezzo, I. Posadas, J. Yu, L. Parente, W. C. Sessa, L. J. Ignarro, and G. Cirino Endothelial nitric oxide synthase activation is critical for vascular leakage during acute inflammation in vivo PNAS, January 18, 2005; 102(3): 904 - 908. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Fulton, M. B. Harris, B. E. Kemp, R. C. Venema, M. B. Marrero, and D. W. Stepp Insulin resistance does not diminish eNOS expression, phosphorylation, or binding to HSP-90 Am J Physiol Heart Circ Physiol, December 1, 2004; 287(6): H2384 - H2393. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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|
|
|
|
 |
|
 P. A. Ortiz, N. J. Hong, and J. L. Garvin Luminal flow induces eNOS activation and translocation in the rat thick ascending limb. II. Role of PI3-kinase and Hsp90 Am J Physiol Renal Physiol, August 1, 2004; 287(2): F281 - F288. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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|
|
|
|
|
|
T. Gidalevitz, C. Biswas, H. Ding, D. Schneidman-Duhovny, H. J. Wolfson, F. Stevens, S. Radford, and Y. Argon Identification of the N-terminal Peptide Binding Site of Glucose-regulated Protein 94 J. Biol. Chem., April 16, 2004; 279(16): 16543 - 16552. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. A. Pritchard Jr., J. Ou, Z. Ou, Y. Shi, J. P. Franciosi, P. Signorino, S. Kaul, C. Ackland-Berglund, K. Witte, S. Holzhauer, et al. Hypoxia-induced acute lung injury in murine models of sickle cell disease Am J Physiol Lung Cell Mol Physiol, April 1, 2004; 286(4): L705 - L714. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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|
|
|
|
|
|
J. Ou, J. T. Fontana, Z. Ou, D. W. Jones, A. W. Ackerman, K. T. Oldham, J. Yu, W. C. Sessa, and K. A. Pritchard Jr. Heat shock protein 90 and tyrosine kinase regulate eNOS NO{middle dot} generation but not NO{middle dot} bioactivity Am J Physiol Heart Circ Physiol, February 1, 2004; 286(2): H561 - H569. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Luo and J. L. Benovic G Protein-coupled Receptor Kinase Interaction with Hsp90 Mediates Kinase Maturation J. Biol. Chem., December 19, 2003; 278(51): 50908 - 50914. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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|
|
|
|
|
|
M. I. Lin, D. Fulton, R. Babbitt, I. Fleming, R. Busse, K. A. Pritchard Jr., and W. C. Sessa Phosphorylation of Threonine 497 in Endothelial Nitric-oxide Synthase Coordinates the Coupling of L-Arginine Metabolism to Efficient Nitric Oxide Production J. Biol. Chem., November 7, 2003; 278(45): 44719 - 44726. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Yoshida and Y. Xia Heat Shock Protein 90 as an Endogenous Protein Enhancer of Inducible Nitric-oxide Synthase J. Biol. Chem., September 19, 2003; 278(38): 36953 - 36958. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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|
|
|
|