Endothelial nitric oxide synthase is segregated from caveolin-1 and localizes to the leading edge of migrating cells.

Abstract:

:The enzyme endothelial Nitric Oxide Synthase (eNOS) is involved in key physiological and pathological processes, including cell motility and apoptosis. It is widely believed that at the cell surface eNOS is localized in caveolae, where caveolin-1 negatively regulates its activity, however, there are still uncertainties on its intracellular distribution. Here, we applied high resolution confocal microscopy to investigate the surface distribution of eNOS in transfected HeLa cells and in human umbilical vein endothelial cells (HUVEC) endogenously expressing the enzyme. In confluent and non-confluent HUVEC and HeLa cells, we failed to detect substantial colocalization between eNOS and caveolin-1 at the cell surface. Instead, in non-confluent cells, eNOS was concentrated in ruffles and at the leading edge of migrating cells, colocalizing with actin filaments and with the raft marker ganglioside G(M1), and well segregated from caveolin-1, which was restricted to the posterior region of the cells. Treatments that disrupted microfilaments caused loss of eNOS from the cell surface and decreased Ca(2+)-stimulated activity, suggesting a role of the cytoskeleton in the localization and function of the enzyme. Our results provide a morphological correlate for the role of eNOS in cell migration and raise questions on the site of interaction between eNOS and caveolin-1.

journal_name

Exp Cell Res

authors

Bulotta S,Cerullo A,Barsacchi R,De Palma C,Rotiroti D,Clementi E,Borgese N

doi

10.1016/j.yexcr.2005.12.014

keywords:

subject

Has Abstract

pub_date

2006-04-01 00:00:00

pages

877-89

issue

6

eissn

0014-4827

issn

1090-2422

pii

S0014-4827(05)00582-3

journal_volume

312

pub_type

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