JCB logo
Avanti Polar Lipids, Inc.
  Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents
Published online 18 September 2000. doi:10.1083/jcb.150.6.1311
This Article
Right arrow Full Text
Right arrow Full Text (PDF, 264K)
Right arrow PPT slides of all figures
Right arrow Alert me when this article is cited
Right arrow Citation Map
Services
Right arrow Email this article
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new content in the JCB
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Higgs, H. N.
Right arrow Articles by Pollard, T. D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Higgs, H. N.
Right arrow Articles by Pollard, T. D.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Facebook   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

© The Rockefeller University Press, 0021-9525/2000//1311 $5.00
The Journal of Cell Biology, Volume 150, Number 6, , 2000 1311-1320

Original Article

Activation by Cdc42 and Pip2 of Wiskott-Aldrich Syndrome Protein (Wasp) Stimulates Actin Nucleation by Arp2/3 Complex



Henry N. Higgsa and Thomas D. Pollarda

a The Salk Institute for Biological Studies, La Jolla, California 92037
The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037.(858) 452-3683(858) 453-4100 x1716

We purified native WASp (Wiskott-Aldrich Syndrome protein) from bovine thymus and studied its ability to stimulate actin nucleation by Arp2/3 complex. WASp alone is inactive in the presence or absence of 0.5 µM GTP-Cdc42. Phosphatidylinositol 4,5 bisphosphate (PIP2) micelles allowed WASp to activate actin nucleation by Arp2/3 complex, and this was further enhanced twofold by GTP-Cdc42. Filaments nucleated by Arp2/3 complex and WASp in the presence of PIP2 and Cdc42 concentrated around lipid micelles and vesicles, providing that Cdc42 was GTP-bound and prenylated. Thus, the high concentration of WASp in neutrophils (9 µM) is dependent on interactions with both acidic lipids and GTP-Cdc42 to activate actin nucleation by Arp2/3 complex. The results also suggest that membrane binding increases the local concentrations of Cdc42 and WASp, favoring their interaction.

Key Words: prenylation • membrane • GBD • neutrophil • thymus

© 2000 The Rockefeller University Press

Abbreviations used in this paper: CF-PE, carboxyfluorescein-labeled PE; GBD, GTPase binding domain; GST, glutathione-S-transferase; HSS, high speed supernatant; PA, phosphatidic acid; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PIP2, phosphatidylinositol 4,5 bisphosphate; PS, phosphatidylserine; PMN, polymorphonuclear leukocytes; WA, WASp/Scar protein COOH-terminal 70–100 amino acids; WASp, Wiskott-Aldrich Syndrome protein.


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Facebook Facebook   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?


This article has been cited by other articles:



  Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents