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

J. Biol. Chem., Vol. 279, Issue 53, 55682-55689, December 31, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/53/55682    most recent M406621200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map 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 Navarro-Lérida, I. Articles by Rodríguez-Crespo, I. Search for Related Content PubMed PubMed Citation Articles by Navarro-Lérida, I. Articles by Rodríguez-Crespo, I. Social Bookmarking                      What's this?

Palmitoylation of Inducible Nitric-oxide Synthase at Cys-3 Is Required for Proper Intracellular Traffic and Nitric Oxide Synthesis^*

Inmaculada Navarro-Lérida, Maria Martha Corvi¶, Alberto Álvarez Barrientos||, Francisco Gavilanes, Luc Gérard Berthiaume**, and Ignacio Rodríguez-Crespo

From the Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas, Universidad Complutense de Madrid, Madrid 28040 Spain, the Department of Cell Biology, MSB-555, University of Alberta, Edmonton, Alberta T6G 2S2, Canada, and the ^||Fundación Centro Nacional de Investigaciones Cardiovasculares, Madrid 28029, Spain

A number of cell types express inducible nitric-oxide synthase (NOS2) in response to exogenous insults such as bacterial lipopolysaccharide or proinflammatory cytokines. Although it has been known for some time that the N-terminal end of NOS2 suffers a post-translational modification, its exact identification has remained elusive. Using radioactive fatty acids, we show herein that NOS2 becomes thioacylated at Cys-3 with palmitic acid. Site-directed mutagenesis of this single residue results in the absence of the radiolabel incorporation. Acylation of NOS2 is completely indispensable for intracellular sorting and ·NO synthesis. In fact, a C3S mutant of NOS2 is completely inactive and accumulates to intracellular membranes that almost totally co-localize with the Golgi marker -cop. Likewise, low concentrations of the palmitoylation blocking agents 2-Br-palmitate or 8-Br-palmitate severely affected the ·NO synthesis of both NOS2 induced in muscular myotubes and transfected NOS2. However, unlike endothelial NOS, palmitoylation of inducible NOS is not involved in its targeting to caveolae. We have created 16 NOS2-GFP chimeras to inspect the effect of the neighboring residues of Cys-3 on the degree of palmitoylation. In this regard, the hydrophobic residue Pro-4 and the basic residue Lys-6 seem to be indispensable for palmitoylation. In addition, agents that block the endoplasmic reticulum to Golgi transit such as brefeldin A and monensin drastically reduced NOS2 activity leading to its accumulation in perinuclear areas. In summary, palmitoylation of NOS2 at Cys-3 is required for both its activity and proper intracellular localization.

Received for publication, June 14, 2004 , and in revised form, October 12, 2004.

^* This work was supported in part by Grants BMC 2003-06631, BMC 2003-05034 (DGICYT), and 08.4/0039.1/2000 (Comunidad Autónoma de Madrid), as well as by CIHR Grant MOP-37955 (to L. G. B.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ Supported by a AHFMR studentship.

^** Alberta Heritage Foundation for Medical Research senior scholar.

To whom correspondence should be addressed. Tel.: 34-91-394-4258; Fax: 34-91-394-4159; E-mail: nacho{at}bbm1.ucm.es.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				A. Martinez-Ruiz and S. Lamas Signalling by NO-induced protein S-nitrosylation and S-glutathionylation: Convergences and divergences Cardiovasc Res, July 15, 2007; 75(2): 220 - 228. [Abstract] [Full Text] [PDF]

				I. Navarro-Lerida, M. Martinez-Moreno, I. Ventoso, A. Alvarez-Barrientos, and I. Rodriguez-Crespo Binding of CAP70 to Inducible Nitric Oxide Synthase and Implications for the Vectorial Release of Nitric Oxide in Polarized Cells Mol. Biol. Cell, July 1, 2007; 18(7): 2768 - 2777. [Abstract] [Full Text] [PDF]

				R. J. Jones, D. Jourd'heuil, J. C. Salerno, S. M. E. Smith, and H. A. Singer iNOS regulation by calcium/calmodulin-dependent protein kinase II in vascular smooth muscle Am J Physiol Heart Circ Physiol, June 1, 2007; 292(6): H2634 - H2642. [Abstract] [Full Text] [PDF]

				I. Navarro-Lerida, A. Alvarez-Barrientos, and I. Rodriguez-Crespo N-terminal palmitoylation within the appropriate amino acid environment conveys on NOS2 the ability to progress along the intracellular sorting pathways J. Cell Sci., April 15, 2006; 119(8): 1558 - 1569. [Abstract] [Full Text] [PDF]

				B. Rotblat, O. Yizhar, R. Haklai, U. Ashery, and Y. Kloog Ras and Its Signals Diffuse through the Cell on Randomly Moving Nanoparticles Cancer Res., February 15, 2006; 66(4): 1974 - 1981. [Abstract] [Full Text] [PDF]

				R. A. Gubitosi-Klug, D. J. Mancuso, and R. W. Gross The human Kv1.1 channel is palmitoylated, modulating voltage sensing: Identification of a palmitoylation consensus sequence PNAS, April 26, 2005; 102(17): 5964 - 5968. [Abstract] [Full Text] [PDF]