|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Papers In Press, published online ahead of print May 15, 2006
Human Microbiology, Tel Aviv University, Sackler School of Medicine, Tel Aviv 69978
Corresponding Author: epick{at}post.tau.ac.il
Activation of the phagocyte NADPH oxidase involves the assembly of a membrane-localized cytochrome b559 with the cytosolic components p47phox, p67phox, p40phox, and the GTPase Rac (1 or 2). In resting phagocytes, Rac is found in the cytosol as a prenylated protein in the GDP-bound form, associated with Rho GDP dissociation inhibitor (RhoGDI). In the process of NADPH oxidase activation, Rac is dissociated from RhoGDI and translocates to the membrane, in concert with the other cytosolic components. The mechanism responsible for dissociation of Rac from RhoGDI is poorly understood. We generated [Rac(1 or 2)-RhoGDI] complexes in vitro from recombinant Rac (1 or 2), prenylated enzymatically, and recombinant RhoGDI, and purified these by anion exchange chromatography. Exposing [Rac(1 or 2)(GDP)-RhoGDI] complexes to liposomes containing four different anionic phospholipids, caused the dissociation of Rac(1 or 2)(GDP) from RhoGDI and its binding to the anionic liposomes. [Rac2(GDP)-RhoGDI] complexes were more resistant to dissociation, reflecting the lesser positive charge of Rac2. Liposomes consisting of neutral phospholipid did not cause dissociation of [Rac(1 or 2)-RhoGDI] complexes. Rac1 exchanged to the hydrolysis-resistant GTP analogue, guanylyl-imidodiphosphate (GMPPNP), associated with RhoGDI with lower affinity than Rac1(GDP) and [Rac1(GMPPNP)-RhoGDI] complexes were more readily dissociated by anionic liposomes. [Rac1(GMPPNP)-RhoGDI] complexes elicited NADPH oxidase activation in native phagocyte membrane liposomes in the presence of p67phox, without the need for an anionic amphiphile, as activator. Both [Rac1(GDP)-RhoGDI] and [Rac1(GMPPNP)-RhoGDI] complexes elicited amphiphile-independent, p67phox-dependent NADPH oxidase activation in phagocyte membrane liposomes enriched in anionic phospholipids but not in membrane liposomes enriched in neutral phospholipids.
J. Biol. Chem, 10.1074/jbc.M600042200
Submitted on January 3, 2006
Revised on May 12, 2006
Accepted on May 15, 2006
Liposomes comprising anionic but not neutral phospholipids cause dissociation of [Rac(1 or 2)-RhoGDI] complexes and support amphiphile-independent NADPH oxidase activation by such complexes
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  Y. Kim, Y.-S. Lee, J. Choe, H. Lee, Y.-M. Kim, and D. Jeoung CD44-Epidermal Growth Factor Receptor Interaction Mediates Hyaluronic Acid-promoted Cell Motility by Activating Protein Kinase C Signaling Involving Akt, Rac1, Phox, Reactive Oxygen Species, Focal Adhesion Kinase, and MMP-2 J. Biol. Chem., August 15, 2008; 283(33): 22513 - 22528. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Ugolev, Y. Berdichevsky, C. Weinbaum, and E. Pick Dissociation of Rac1(GDP){middle dot}RhoGDI Complexes by the Cooperative Action of Anionic Liposomes Containing Phosphatidylinositol 3,4,5-Trisphosphate, Rac Guanine Nucleotide Exchange Factor, and GTP J. Biol. Chem., August 8, 2008; 283(32): 22257 - 22271. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. C. Chae, J. H. Kim, K. L. Kim, H. W. Kim, H. Y. Lee, W. D. Heo, T. Meyer, P.-G. Suh, and S. H. Ryu Phospholipase D Activity Regulates Integrin-mediated Cell Spreading and Migration by Inducing GTP-Rac Translocation to the Plasma Membrane Mol. Biol. Cell, July 1, 2008; 19(7): 3111 - 3123. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Chianale, S. Cutrupi, E. Rainero, G. Baldanzi, P. E. Porporato, S. Traini, N. Filigheddu, V. F. Gnocchi, M. M. Santoro, O. Parolini, et al. Diacylglycerol Kinase-{alpha} Mediates Hepatocyte Growth Factor-induced Epithelial Cell Scatter by Regulating Rac Activation and Membrane Ruffling Mol. Biol. Cell, December 1, 2007; 18(12): 4859 - 4871. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Berdichevsky, A. Mizrahi, Y. Ugolev, S. Molshanski-Mor, and E. Pick Tripartite Chimeras Comprising Functional Domains Derived from the Cytosolic NADPH Oxidase Components p47phox, p67phox, and Rac1 Elicit Activator-independent Superoxide Production by Phagocyte Membranes: AN ESSENTIAL ROLE FOR ANIONIC MEMBRANE PHOSPHOLIPIDS J. Biol. Chem., July 27, 2007; 282(30): 22122 - 22139. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M.-H. Paclet, S. Berthier, L. Kuhn, J. Garin, and F. Morel Regulation of phagocyte NADPH oxidase activity: identification of two cytochrome b558 activation states FASEB J, April 1, 2007; 21(4): 1244 - 1255. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. McDonald, R. Veluthakal, H. Kaur, and A. Kowluru Biologically active lipids promote trafficking and membrane association of Rac1 in insulin-secreting INS 832/13 cells Am J Physiol Cell Physiol, March 1, 2007; 292(3): C1216 - C1220. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. DerMardirossian, G. Rocklin, J.-Y. Seo, and G. M. Bokoch Phosphorylation of RhoGDI by Src Regulates Rho GTPase Binding and Cytosol-Membrane Cycling Mol. Biol. Cell, November 1, 2006; 17(11): 4760 - 4768. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |