HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: fj.06-7088comv1 21/11/2655    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Joshi, M. S. Articles by Bauer, J. A. Search for Related Content PubMed PubMed Citation Articles by Joshi, M. S. Articles by Bauer, J. A.

Biochemical consequences of the NOS3 Glu298Asp variation in human endothelium: altered caveolar localization and impaired response to shear

Mandar S. Joshi^*, Chieko Mineo, Philip W. Shaul and John Anthony Bauer^*,1

^* Center for Cardiovascular Medicine, Columbus Children's Research Institute, Columbus, Ohio, USA; and

University of Texas Southwestern Medical Center at Dallas, Department of Pediatrics, Dallas, Texas, USA

¹Correspondence: Center for Cardiovascular Medicine, Columbus Children's Research Institute, 700 Children's Dr., Columbus, OH 43205, USA. E-mail: bauerj{at}ccri.net

Human endothelial nitric oxide synthase (NOS3) gene polymorphism at Exon 7 (Glu298Asp) has been linked to vascular endothelial dysfunction, but the mechanisms are not defined. Shear is a key modulator of NOS3 function in vivo and association with caveolae is important for the control of NOS3 protein activity. Here we tested the hypothesis that altered enrichment of NOS3 in the caveolar membrane defines Glu298Asp genotype-specific responses and NOS3 activity. Basal caveolar membrane enrichment was carried out to quantitate the NOS3 enrichment in caveolae. Cells were subjected to shear and NOS3 protein levels, phosphorylation, enzyme function were investigated. Variant genotypes had lower NOx production pre- and post-shear, but no genotype-dependent alterations in pNOS3 were observed. Asp variants had significantly lower NOS3 enrichment in the caveolar membrane fraction. Further, immunoprecipitation studies demonstrated that Asp variants had substantially less NOS3/Cav-1 association (40%) during static conditions. Furthermore, acute shear causes impaired NOS3/Cav-1 dissociation in Asp variants. The results from immunoprecipitation studies were in complete agreement with caveolar membrane preparation findings. Collectively, these data demonstrate functional consequences of the Glu298Asp NOS3 variation and further define disruption of NOS3 caveolar localization and shear-induced mobilization as the primary mechanism responsible for these differences.—Joshi, M. S., Mineo, C., Shaul, P. W., Bauer, J. A. Biochemical consequences of the NOS3 Glu298Asp variation in human endothelium: altered caveolar localization and impaired response to shear.

Key Words: caveolin-1 • endothelial nitric oxide synthase • functional genomics • gene polymorphism • protein-protein interactions

This article has been cited by other articles:

				A. Imamura, R. Takahashi, R. Murakami, H. Kataoka, X. W. Cheng, Y. Numaguchi, T. Murohara, and K. Okumura The effects of endothelial nitric oxide synthase gene polymorphisms on endothelial function and metabolic risk factors in healthy subjects: the significance of plasma adiponectin levels Eur. J. Endocrinol., February 1, 2008; 158(2): 189 - 195. [Abstract] [Full Text] [PDF]