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

J. Biol. Chem., Vol. 282, Issue 4, 2567-2575, January 26, 2007

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: 282/4/2567    most recent M604560200v1 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 Mellgren, R. L. Articles by McNeil, P. L. Search for Related Content PubMed PubMed Citation Articles by Mellgren, R. L. Articles by McNeil, P. L. Social Bookmarking                      What's this?

Calpain Is Required for the Rapid, Calcium-dependent Repair of Wounded Plasma Membrane^*

Ronald L. Mellgren¹, Wenli Zhang, Katsuya Miyake, and Paul L. McNeil^¶

From the Department of Physiology, Pharmacology, Metabolism, and Cardiovascular Sciences, University of Toledo College of Medicine, Toledo, Ohio 43614, the Institute of Molecular Medicine and Genetics, and the ^¶Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta, Georgia 30912

Mammalian cells require extracellular calcium ion to undergo rapid plasma membrane repair seconds after mechanical damage. Utilizing transformed fibroblasts from calpain small subunit knock-out (Capns1^-/-) mouse embryos, we now show that the heterodimeric, typical subclass of calpains is required for calcium-mediated survival after plasma membrane damage caused by scraping a cell monolayer. Survival of scrape-damaged Capns1^-/- cells was unaffected by calcium in the scraping medium, whereas more Capns1^+/+ cells survived when calcium was present. Calcium-mediated survival was increased when Capns1^-/- cells were scraped in the presence of purified m- or µ-calpain. Survival rates of scraped Capns1^+/+, HFL-1, or Chinese hamster ovary cells were decreased by the calpain inhibitor, calpeptin, or the highly specific calpain inhibitor protein, calpastatin. Capns1^-/- cells failed to reseal following laser-induced membrane disruption, demonstrating that their decreased survival after scraping resulted, at least in part, from failed membrane repair. Proteomic and immunologic analyses demonstrated that the known calpain substrates talin and vimentin were exposed at the cell surface and processed by calpain following cell scraping. Autoproteolytic activation of calpain at the scrape site was evident at the earliest time point analyzed and appeared to precede proteolysis of talin and vimentin. The results indicate that conventional calpains are required for calcium-facilitated survival after plasma membrane damage and may act by localized remodeling of the cortical cytoskeleton at the injury site.

Received for publication, May 12, 2006 , and in revised form, October 17, 2006.

^* This work was supported in part by awards from the American Heart Association (RLM), and the National Aeronautics and Space Administration (PLM). 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 supplemental Figs. S1 and S2 and videos 1 and 2.

¹ To whom correspondence should be addressed: Dept. of Physiology, Pharmacology, Metabolism, and Cardiovascular Sciences, Mail Stop 1008, Health Science Campus, University of Toledo College of Medicine, 3000 Arlington Ave., Toledo, OH 43614-2598. Tel.: 419-383-4182; Fax: 419-383-2871; E-mail: rmellgren{at}meduohio.edu.

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

This article has been cited by other articles:

				T. Yamazaki, Y. Kawamura, A. Minami, and M. Uemura Calcium-Dependent Freezing Tolerance in Arabidopsis Involves Membrane Resealing via Synaptotagmin SYT1 PLANT CELL, December 1, 2008; 20(12): 3389 - 3404. [Abstract] [Full Text] [PDF]

				Y. Huang, A. de Morree, A. van Remoortere, K. Bushby, R. R. Frants, J. T. Dunnen, and S. M. van der Maarel Calpain 3 is a modulator of the dysferlin protein complex in skeletal muscle Hum. Mol. Genet., June 15, 2008; 17(12): 1855 - 1866. [Abstract] [Full Text] [PDF]

				R. L. Mellgren and X. Huang Fetuin A Stabilizes m-Calpain and Facilitates Plasma Membrane Repair J. Biol. Chem., December 7, 2007; 282(49): 35868 - 35877. [Abstract] [Full Text] [PDF]