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

J. Biol. Chem., Vol. 279, Issue 23, 24460-24466, June 4, 2004

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: 279/23/24460    most recent M312764200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted 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 Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Chodniewicz, D. Articles by Zhelev, D. V. Search for Related Content PubMed PubMed Citation Articles by Chodniewicz, D. Articles by Zhelev, D. V. Social Bookmarking                      What's this?

Experimental Evidence for the Limiting Role of Enzymatic Reactions in Chemoattractant-induced Pseudopod Extension in Human Neutrophils^*

David Chodniewicz, Abdullatif M. Alteraifi¶, and Doncho V. Zhelev||

From the Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708-0300 and ^¶Department of Mechanical Engineering, United Arab Emirates University, Al Ain, United Arab Emirates

Chemoattractant-stimulated pseudopod growth in human neutrophils was used as a model system to study the rate-limiting mechanism of cytoskeleton rearrangement induced by activated G-protein-coupled receptors. Cells were activated with N-formyl-Met-Leu-Phe, and the temperature dependence of the rate of pseudopod extension was measured in the presence of pharmacological inhibitors with known mechanisms of action. Three groups of inhibitors were used: (i) inhibitors sequestering substrates involved in F-actin polymerization (latrunculin A for G-actin and cytochalasin D for actin filament-free barbed ends) or sequestering secondary messengers (PIP-binding peptide for phosphoinositide lipids); (ii) competitively binding inhibitors (Akt-inhibitor for Akt/protein kinase B); and (iii) inhibitors that reduce enzyme activity (wortmannin for phosphoinositide 3-kinase and chelerythrine for protein kinase C). The experimental data are consistent with a model in which the relative involvement of a given pathway of F-actin polymerization to the measured rate of pseudopod extension is limited by a slowest (bottleneck) reaction in the cascade of reactions involved in the overall signaling pathway. The approach we developed was used to demonstrate that chemoattractant-induced pseudopod growth and mechanically stimulated cytoskeleton rearrangement are controlled by distinct pathways of F-actin polymerization.

Received for publication, November 21, 2003 , and in revised form, March 19, 2004.

^* This work was supported by Grants HL57629 and NCBC ARG-0031 (to D. V. Z.). Blood drawing was supported by National Institutes of Health Grant M01-RR-30 (to the General Clinical Research Centers Program at Duke University). 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.

The on-line version of this article (available at http://www.jbc.org) contains one table and two figures.

Recipient of a fellowship from National Institutes of Health Research Training Grant GM08555. Present address: Department of Immunology, The Scripps Research Institute, La Jolla, CA 92037.

^|| To whom correspondence should be addressed: Dept. of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708-0300. Tel.: 919-660-5335; Fax: 919-660-8963; E-mail: dvzh{at}duke.edu.

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

This article has been cited by other articles:

				M.-P. Belot, L. Abdennebi-Najar, F. Gaudin, M. Lieberherr, V. Godot, J. Taieb, D. Emilie, and V. Machelon Progesterone reduces the migration of mast cells toward the chemokine stromal cell-derived factor-1/CXCL12 with an accompanying decrease in CXCR4 receptors Am J Physiol Endocrinol Metab, May 1, 2007; 292(5): E1410 - E1417. [Abstract] [Full Text] [PDF]