QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

First published online 22 July 2003
doi: 10.1242/jcs.00664

This Article Figures Only Full Text Full Text (PDF) All Versions of this Article: jcs.00664v1 116/17/3645    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 Chatterjee, S. Articles by Shah, V. Search for Related Content PubMed PubMed Citation Articles by Chatterjee, S. Articles by Shah, V.

Inhibition of GTP-dependent vesicle trafficking impairs internalization of plasmalemmal eNOS and cellular nitric oxide production

¹ GI Research Unit, Department of Physiology, and Tumor Biology Program, Mayo Clinic, Rochester, MN 55905, USA
² Division of Cardiovascular Disease, and Molecular Medicine Program, Mayo Clinic, Rochester, MN 55905, USA

^* Author for correspondence (e-mail: shah.vijay{at}mayo.edu)

Accepted 12 May 2003

The Ca²⁺ mobilizing peptide, bradykinin (BK), stimulates endothelial nitric oxide synthase (eNOS)-derived cellular nitric oxide (NO) production in association with altering the subcellular distribution of the enzyme. In the present study we examine the influence of cellular GTPases, particularly the large GTPase dynamin, on BK-mediated eNOS localization and cellular NO production. BK stimulation of ECV cells, which were stably transfected with eNOS-GFP (eNOS-GFP ECV304), increased NO production. This was associated with the mobilization of eNOS-GFP protein into Triton X-100-insoluble fractions of cell lysates, and an internalization of plasmalemmal eNOS-GFP in live and fixed ECV 304 cells. Incubation of digitonin-permeabilized ECV304 cells with the non-hydrolyzed GTP analog, GTP--S, abrogated the BK-mediated internalization of eNOS-GFP as assessed by confocal microscopy. Conversely, inhibition of clathrin-dependent endocytosis, via overexpression of AP 180 or pretreatment of cells with chlorpromazine, did not influence BK-mediated eNOS redistribution. Furthermore, specific inhibition of dynamin-2 GTPase function by overexpression of a dominant negative construct, K44A, prevented the BK-mediated enrichment of eNOS-GFP within low buoyant density, caveolin-enriched fractions of eNOS-GFP ECV304 cell lysates. Dynamin-2 K44A overexpression also markedly impaired BK-dependent, L-NAME-inhibited NO production as did incubation of permeabilized cells with GTP--s. These studies demonstrate that disruption of dynamin- and GTP-dependent, but clathrin-independent, vesicle trafficking pathways impairs BK-dependent cellular NO production, via inhibition of the internalization of eNOS-containing plasmalemmal vesicles.

Key words: Nitric oxide, Nitric oxide synthase, Bradykinin, Dynamin-2

This article has been cited by other articles:

				F. A. Sanchez, D. D. Kim, R. G. Duran, C. J. Meininger, and W. N. Duran Internalization of eNOS via caveolae regulates PAF-induced inflammatory hyperpermeability to macromolecules Am J Physiol Heart Circ Physiol, October 1, 2008; 295(4): H1642 - H1648. [Abstract] [Full Text] [PDF]

				N. Kang-Decker, S. Cao, S. Chatterjee, J. Yao, L. J. Egan, D. Semela, D. Mukhopadhyay, and V. Shah Nitric oxide promotes endothelial cell survival signaling through S-nitrosylation and activation of dynamin-2 J. Cell Sci., February 1, 2007; 120(3): 492 - 501. [Abstract] [Full Text] [PDF]

				N. A. Maniatis, V. Brovkovych, S. E. Allen, T. A. John, A. N. Shajahan, C. Tiruppathi, S. M. Vogel, R. A. Skidgel, A. B. Malik, and R. D. Minshall Novel Mechanism of Endothelial Nitric Oxide Synthase Activation Mediated by Caveolae Internalization in Endothelial Cells Circ. Res., October 13, 2006; 99(8): 870 - 877. [Abstract] [Full Text] [PDF]

				S. Mukherjee, M. Tessema, and A. Wandinger-Ness Vesicular Trafficking of Tyrosine Kinase Receptors and Associated Proteins in the Regulation of Signaling and Vascular Function Circ. Res., March 31, 2006; 98(6): 743 - 756. [Abstract] [Full Text] [PDF]

				A. Icking, S. Matt, N. Opitz, A. Wiesenthal, W. Muller-Esterl, and K. Schilling NOSTRIN functions as a homotrimeric adaptor protein facilitating internalization of eNOS J. Cell Sci., November 1, 2005; 118(21): 5059 - 5069. [Abstract] [Full Text] [PDF]

				M. Schleicher, F. Brundin, S. Gross, W. Muller-Esterl, and S. Oess Cell Cycle-Regulated Inactivation of Endothelial NO Synthase through NOSIP-Dependent Targeting to the Cytoskeleton Mol. Cell. Biol., September 15, 2005; 25(18): 8251 - 8258. [Abstract] [Full Text] [PDF]

				J. S. Lee, N. Kang Decker, S. Chatterjee, J. Yao, S. Friedman, and V. Shah Mechanisms of Nitric Oxide Interplay with Rho GTPase Family Members in Modulation of Actin Membrane Dynamics in Pericytes and Fibroblasts Am. J. Pathol., June 1, 2005; 166(6): 1861 - 1870. [Abstract] [Full Text] [PDF]

				I. Lim, S. J Gibbons, G. L. Lyford, S. M. Miller, P. R. Strege, M. G. Sarr, S. Chatterjee, J. H. Szurszewski, V. H. Shah, and G. Farrugia Carbon monoxide activates human intestinal smooth muscle L-type Ca2+ channels through a nitric oxide-dependent mechanism Am J Physiol Gastrointest Liver Physiol, January 1, 2005; 288(1): G7 - G14. [Abstract] [Full Text] [PDF]

				J.-P. Gratton, P. Bernatchez, and W. C. Sessa Caveolae and Caveolins in the Cardiovascular System Circ. Res., June 11, 2004; 94(11): 1408 - 1417. [Abstract] [Full Text] [PDF]