{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T12:21:12Z","timestamp":1790857272740,"version":"4.1.0"},"reference-count":40,"publisher":"The Company of Biologists","issue":"17","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2003,9,1]]},"abstract":"<jats:p>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-\u03b3-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-\u03b3-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.<\/jats:p>","DOI":"10.1242\/jcs.00664","type":"journal-article","created":{"date-parts":[[2003,7,29]],"date-time":"2003-07-29T00:26:32Z","timestamp":1059438392000},"page":"3645-3655","source":"Crossref","is-referenced-by-count":42,"title":["Inhibition of GTP-dependent vesicle trafficking impairs internalization of plasmalemmal eNOS and cellular nitric oxide production"],"prefix":"10.1242","volume":"116","author":[{"given":"Suvro","family":"Chatterjee","sequence":"first","affiliation":[{"name":"GI Research Unit, Department of Physiology, and Tumor Biology Program, Mayo Clinic, Rochester, MN 55905, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sheng","family":"Cao","sequence":"additional","affiliation":[{"name":"GI Research Unit, Department of Physiology, and Tumor Biology Program, Mayo Clinic, Rochester, MN 55905, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Timothy E.","family":"Peterson","sequence":"additional","affiliation":[{"name":"Division of Cardiovascular Disease, and Molecular Medicine Program, Mayo Clinic, Rochester, MN 55905, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert D.","family":"Simari","sequence":"additional","affiliation":[{"name":"Division of Cardiovascular Disease, and Molecular Medicine Program, Mayo Clinic, Rochester, MN 55905, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vijay","family":"Shah","sequence":"additional","affiliation":[{"name":"GI Research Unit, Department of Physiology, and Tumor Biology Program, Mayo Clinic, Rochester, MN 55905, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"237","reference":[{"key":"2021042517390797300_REF1","doi-asserted-by":"crossref","unstructured":"Bernier, S., Haldar, S. and Michel, T. (2000). Bradykinin-regulated interactions of the mitogen-activated protein kinase pathway with the endothelial nitric-oxide synthase. J. Biol. Chem.275,30707-30715.","DOI":"10.1074\/jbc.M005116200"},{"key":"2021042517390797300_REF2","doi-asserted-by":"crossref","unstructured":"Blair, A., Shaul, P., Yuhanna, I., Conrad, P. and Smart, E.(1999). Oxidized low density lipoprotein displaces endothelial nitric-oxide synthase (eNOS) from plasmalemmel caveolae and impairs eNOS activation. J. Biol. Chem.274,32512-32519.","DOI":"10.1074\/jbc.274.45.32512"},{"key":"2021042517390797300_REF3","doi-asserted-by":"crossref","unstructured":"Brown, J., Reading, S., Jones, S., Fitchett, C., Howl, J.,Martin, A., Longland, C., Michelangeli, F., Dubrova, Y. and Brown, C.(2000). Critical evaluation of ECV304 as a human endothelial cell model defined by genetic analysis and functional responses: a comparison with the human bladder cancer derived epithelial cell line T24\/83. Lab. Invest.80,37-45.","DOI":"10.1038\/labinvest.3780006"},{"key":"2021042517390797300_REF4","doi-asserted-by":"crossref","unstructured":"Cao, H., Thompson, H., Krueger, E. and McNiven, M.(2000). Disruption of Golgi structure and function in mammalian cells expressing mutant dynamin. J. Cell Sci.113,1993-2002.","DOI":"10.1242\/jcs.113.11.1993"},{"key":"2021042517390797300_REF5","doi-asserted-by":"crossref","unstructured":"Cao, S., Yao, J. and Shah, V. (2003) The proline-rich domain of dynamin-2 is responsible for dynamin-dependent in vitro potentiation of eNOS activity via selective effects on reductase domain function. J. Biol. Chem.278,5894-5901.","DOI":"10.1074\/jbc.M212546200"},{"key":"2021042517390797300_REF6","doi-asserted-by":"crossref","unstructured":"Cao, S., Yao, Y., McCabe, T., Yao, Q., Katusic, Z., Sessa, W. and Shah, V. (2001). Direct interaction between eNOS and dynamin-2:Implications for NOS function. J. Biol. Chem.276,14249-14256.","DOI":"10.1074\/jbc.M006258200"},{"key":"2021042517390797300_REF7","doi-asserted-by":"crossref","unstructured":"Chavrier, P. and Goud, B. (1999). The role of ARF and Rab GTPase in membrane transport. Curr. Opin. Cell Biol.11,466-475.","DOI":"10.1016\/S0955-0674(99)80067-2"},{"key":"2021042517390797300_REF8","doi-asserted-by":"crossref","unstructured":"Dessy, C., Kelly, R., Balligand, J.-L. and Feron, O.(2000). Dynamin mediates caveolar sequestration of muscarinic cholinergic receptors and alteration in NO signaling. EMBO J.19,4272-4280.","DOI":"10.1093\/emboj\/19.16.4272"},{"key":"2021042517390797300_REF9","doi-asserted-by":"crossref","unstructured":"Ford, M. G., Pearse, B. M., Higgins, M. K., Vallis, Y., Owen, D. J., Gibson, A., Hopkins, C. R., Evans, P. R. and McMahon, H. T.(2001). AP 180 recruits clathrin to membranes: structure of the ANTH domain bound to PtdIns (4,5)P2. Science9,1051-1055.","DOI":"10.1126\/science.291.5506.1051"},{"key":"2021042517390797300_REF10","doi-asserted-by":"crossref","unstructured":"Fulton, D., Fontana, J., Sowa, G., Gratton, J.-P., Lin, M., Li,K.-X., Michell, B., Kemp, B., Rodman, D. and Sessa, W.(2002). Localization of endothelial nitric-oxide synthase phosphorylated on serine 1179 ane nitric oxide on Golgi and plasma membrane defines the existence of two pools of active enzymes. J. Biol. Chem.277,4277-4284.","DOI":"10.1074\/jbc.M106302200"},{"key":"2021042517390797300_REF11","doi-asserted-by":"crossref","unstructured":"Goetz, R., Thatte, H., Prabhakar, P., Cho, M., Michel, T. and Golan, D. (1999). Estradiol induces the calcium-dependent translocation of endothelial nitric oxide synthase. Proc. Natl. Acad. Sci. USA96,2788-2793.","DOI":"10.1073\/pnas.96.6.2788"},{"key":"2021042517390797300_REF12","doi-asserted-by":"crossref","unstructured":"Harris, M., Ju, H., Venema, V., Liang, H., Zou, R., Michell, B.,Chen, Z., Kemp, B. and Venema, R. (2001). Reciprocal phosphorylation and regulation of endothelial nitric-oxide synthase in response to bradykinin stimulation. J. Biol. Chem.276,16587-16591.","DOI":"10.1074\/jbc.M100229200"},{"key":"2021042517390797300_REF13","doi-asserted-by":"crossref","unstructured":"Henley, J., Krueger, E., Oswald, B. and McNiven, M.(1998). Dynamin-mediated internalization of caveolae. J. Cell Biol.141,85-99.","DOI":"10.1083\/jcb.141.1.85"},{"key":"2021042517390797300_REF14","doi-asserted-by":"crossref","unstructured":"Kojima, H., Nakatsubo, N., Kikuchi, K., Sawahara, S., Kirino,Y., Nagoshi, H., Hirata, Y. and Nagano, T. (1998). Detection and imaging of nitric oxide with novel fluorescent indicators:diaminofluoresceins. Annal. Chem.70,2446-2453.","DOI":"10.1021\/ac9801723"},{"key":"2021042517390797300_REF15","doi-asserted-by":"crossref","unstructured":"Liu, J., Garcia-Cardena, G. and Sessa, W.(1995). Biosynthesis and palmitoylation of endothelial nitric oxide synthase: mutagenesis of palmitoylation sites, cysteine-15 and\/or -26,argues against depalmitoylation-induced translocation of the enzyme. Biochemistry34,12333-12340.","DOI":"10.1021\/bi00038a029"},{"key":"2021042517390797300_REF16","doi-asserted-by":"crossref","unstructured":"McNiven, M. (1998). Dynamin: a molecular motor with pinchase action. Cell94,151-154.","DOI":"10.1016\/S0092-8674(00)81414-2"},{"key":"2021042517390797300_REF17","doi-asserted-by":"crossref","unstructured":"Michel, T., Li, G. K. and Busconi, L. (1993). Phosphorylation and subcellular translocation of endothelial nitric oxide synthase. Proc Natl. Acad. Sci. USA90,6252-6256.","DOI":"10.1073\/pnas.90.13.6252"},{"key":"2021042517390797300_REF18","doi-asserted-by":"crossref","unstructured":"Mobasheri, A., Errington, R. J., Golding, S., Hall, A. C. and Urban, P. G. (1997). Characterization of the Na+,K+-ATPase in isolated bovine articular chondrocytes; molecular evidence for multiple and isoforms. Cell Biol. Int.21,201-212.","DOI":"10.1006\/cbir.1997.0137"},{"key":"2021042517390797300_REF19","doi-asserted-by":"crossref","unstructured":"Oh, P., McIntosh, P. and Schnitzer, J. E.(1998). Dynamin at the neck of caveolae mediates their budding to form transport vesicles by GTP-driven fission from the plasma membrane of endothelium. J. Cell Biol.141,101-114.","DOI":"10.1083\/jcb.141.1.101"},{"key":"2021042517390797300_REF20","doi-asserted-by":"crossref","unstructured":"Palmer, R., Ferrige, A. and Moncada, S. (1987). Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor. Nature327,524-526.","DOI":"10.1038\/327524a0"},{"key":"2021042517390797300_REF21","doi-asserted-by":"crossref","unstructured":"Papapetropoulos, A., Rudic, R. and Sessa, W.(1999). Molecular control of nitric oxide synthases in the cardiovascular system. Cardiovasc. Res.43,509-520.","DOI":"10.1016\/S0008-6363(99)00161-3"},{"key":"2021042517390797300_REF22","doi-asserted-by":"crossref","unstructured":"Paxinou, E., Weisse, M., Chen, Q., Souza, J., Hertkorn, C.,Selak, M., Daikhin, E., Yudkoff, M., Sowa, G., Sessa, W. et al.(2001). Dynamic regulation of metabolism and respiration by endogenously produced nitric oxide protects against oxidative stress. Proc. Natl. Acad. Sci. USA98,11575-11580.","DOI":"10.1073\/pnas.201293198"},{"key":"2021042517390797300_REF23","doi-asserted-by":"crossref","unstructured":"Peterson, T., Poppa, V., Ueba, H., Wu, A., Yan, C. and Berk,B. (1999). Opposing effects of reactive oxygen species and cholesterol on endothelial nitric oxide synthase and endothelial cell caveolae. Circ. Res.85,29-37.","DOI":"10.1161\/01.RES.85.1.29"},{"key":"2021042517390797300_REF24","unstructured":"Petris, M. J., Smith, K., Lee, J. and Thiele, D. J.(2002). J. Biol Chem.278,9639-9646."},{"key":"2021042517390797300_REF25","unstructured":"Pierce, K., Maudsley, S., Daaka, Y., Luttrell, L. and Lefkowitz,R. (1999). Role of endocytosis in the activation of the ERK cascade by sequestering and nonsequestering G protein-coupled receptors. Proc. Natl. Acad. Sci. USA97,1489-1494."},{"key":"2021042517390797300_REF26","doi-asserted-by":"crossref","unstructured":"Prabhakar, P., Cheng, V. and Michel, T. (2000). A chimeric transmembrane domain directs endothelial nitric-oxide synthase palmitoylation and targeting to plasmalemmel caveolae. J. Biol. Chem.275,19416-19421.","DOI":"10.1074\/jbc.M001952200"},{"key":"2021042517390797300_REF27","doi-asserted-by":"crossref","unstructured":"Prabhakar, P., Thatte, H., Goetz, R., Cho, M., Golan, D. and Michel, T. (1998). Receptor-regulated translocation of endothelial nitric-oxide synthase. J. Biol. Chem.273,27383-27388.","DOI":"10.1074\/jbc.273.42.27383"},{"key":"2021042517390797300_REF28","doi-asserted-by":"crossref","unstructured":"Rudic, R. (1998). Direct evidence for the importance of endothelium-derived nitric oxide in vascular remodeling. J. Clin. Invest.101,731-736.","DOI":"10.1172\/JCI1699"},{"key":"2021042517390797300_REF29","doi-asserted-by":"crossref","unstructured":"Schekman, R. and Orci, L. (1996). Coat proteins and vesicle budding. Science271,1526-1533.","DOI":"10.1126\/science.271.5255.1526"},{"key":"2021042517390797300_REF30","doi-asserted-by":"crossref","unstructured":"Schnitzer, J., Oh, P. and McIntosh, D. (1996). Role of GTP hydrolysis in fission of caveolae directly from plasma membranes. Science274,239-242.","DOI":"10.1126\/science.274.5285.239"},{"key":"2021042517390797300_REF31","doi-asserted-by":"crossref","unstructured":"Sessa, W. C., Garcia-Cardena, G., Liu, J., Keh, A., Pollock, J. S., Bradley, J., Thiru, S., Braverman, I. M. and Desai, K. M.(1995). The Golgi association of endothelial nitric oxide synthase is necessary for the efficient synthesis of nitric oxide. J. Biol. Chem.270,17641-17644.","DOI":"10.1074\/jbc.270.30.17641"},{"key":"2021042517390797300_REF32","doi-asserted-by":"crossref","unstructured":"Shah, V., Haddad, F., Garcia-Cardena, G., Frangos, J., Mennone,A., Groszmann, R. and Sessa, W. (1997). Liver sinusoidal endothelial cells are responsible for nitric oxide modulation of hepatic resistance. J. Clin. Invest.100,2923-2930.","DOI":"10.1172\/JCI119842"},{"key":"2021042517390797300_REF33","doi-asserted-by":"crossref","unstructured":"Shah, V., Toruner, M., Haddad, F., Cadelina, G.,Papapetropoulos, A., Sessa, W. and Groszmann, R. (1999). Impaired endothelial nitric oxide synthase activity associated with enhanced caveolin binding in experimental liver cirrhosis. Gastroenterology117,1222-1228.","DOI":"10.1016\/S0016-5085(99)70408-7"},{"key":"2021042517390797300_REF34","doi-asserted-by":"crossref","unstructured":"Shaul, P. (2002). Regulation of endothelial nitric oxide synthase: location, location, location. Annu. Rev. Physiol.64,749-774.","DOI":"10.1146\/annurev.physiol.64.081501.155952"},{"key":"2021042517390797300_REF35","doi-asserted-by":"crossref","unstructured":"Shaul, P., Smart, E., Robinson, L., German, Z., Yuhanna, I.,Ying, Y., Anderson, R. and Michel, T. (1996). Acylation targets endothelial nitric-oxide synthase to plasmalemmal caveolae. J. Biol. Chem.271,6518-6522.","DOI":"10.1074\/jbc.271.11.6518"},{"key":"2021042517390797300_REF36","doi-asserted-by":"crossref","unstructured":"Smart, E., Ying, Y., Mineo, C. and Anderson, R.(1995). A detergent-free method for purifying caveolae membrane from tissue cultured cells. Proc. Natl. Acad. Sci. USA92,10104-10108.","DOI":"10.1073\/pnas.92.22.10104"},{"key":"2021042517390797300_REF37","doi-asserted-by":"crossref","unstructured":"Sowa, G., Liu, J., Papapetropoulos, A., Rex-Haffner, M., Hughes,T. and Sessa, W. (1999). Trafficking of endothelial nitric-oxide synthase in living cells. J. Biol. Chem.274,22524-22531.","DOI":"10.1074\/jbc.274.32.22524"},{"key":"2021042517390797300_REF38","doi-asserted-by":"crossref","unstructured":"Thuringer, D., Maulon, L. and Frelin, C.(2002). Rapid transaction of the vascular endothelial growth factor receptor KDR\/Flk-1 by the bradykinin B2 receptor contributes to endothelial nitric-oxide synthase activation in cardiac capillary endothelial cells. J. Biol. Chem.277,2028-2032.","DOI":"10.1074\/jbc.M109493200"},{"key":"2021042517390797300_REF39","doi-asserted-by":"crossref","unstructured":"Venema, V., Marrero, M. and Venema, R. (1996). Bradykinin-stimulated protein tyrosine phosphorylation promotes endothelial nitric oxide synthase translocation to the cytoskeleton. Biochem. Biophys. Res. Commun.226,703-710.","DOI":"10.1006\/bbrc.1996.1417"},{"key":"2021042517390797300_REF40","doi-asserted-by":"crossref","unstructured":"Zimmerman, K., Opitz, N., Dedio, J., Renne, C., Muller-Esterl,W. and Oess, S. (2002). NOSTRIN: A protein modulating nitric oxide production and subcellular distribution of eNOS Proc. Natl. Acad. Sci. USA99,17167-17172.","DOI":"10.1073\/pnas.252345399"}],"container-title":["Journal of Cell Science"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/journals.biologists.com\/jcs\/article-pdf\/116\/17\/3645\/1362324\/3645.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"http:\/\/journals.biologists.com\/jcs\/article-pdf\/116\/17\/3645\/1362324\/3645.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,4,25]],"date-time":"2021-04-25T17:51:44Z","timestamp":1619373104000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.biologists.com\/jcs\/article\/116\/17\/3645\/27146\/Inhibition-of-GTP-dependent-vesicle-trafficking"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2003,9,1]]},"references-count":40,"journal-issue":{"issue":"17","published-print":{"date-parts":[[2003,9,1]]}},"URL":"https:\/\/doi.org\/10.1242\/jcs.00664","relation":{},"ISSN":["1477-9137","0021-9533"],"issn-type":[{"value":"1477-9137","type":"electronic"},{"value":"0021-9533","type":"print"}],"subject":[],"published":{"date-parts":[[2003,9,1]]}}}