QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

First published online 1 August 2006
doi: 10.1242/jcs.03089

This Article Figures Only Full Text Full Text (PDF) All Versions of this Article: jcs.03089v1 119/16/3435    most recent Alert me when this article is cited Alert me if a correction is posted 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 Lampe, P. D. Articles by Burt, J. M. Search for Related Content PubMed PubMed Citation Articles by Lampe, P. D. Articles by Burt, J. M. Social Bookmarking                         What's this?

Analysis of Connexin43 phosphorylated at S325, S328 and S330 in normoxic and ischemic heart

Paul D. Lampe^1,*, Cynthia D. Cooper¹, Timothy J. King^1, and Janis M. Burt²

¹ Molecular Diagnostics Program, Fred Hutchinson Cancer Research Center and Department of Pathobiology, University of Washington, 1100 Fairview Avenue N., M5C800, P.O. Box 19024, Seattle, WA 98109, USA
² Department of Physiology, University of Arizona, Tucson, AZ

^* Author for correspondence (e-mail: plampe{at}fhcrc.org)

Accepted 7 June 2006

The functional consequences of Connexin43 (Cx43) phosphorylation remain largely unexplored. Using an antibody that specifically recognizes Cx43 phosphorylated at serine residues 325, 328 and/or 330 (pS325/328/330-Cx43), we show that labeling of this form of Cx43 as well as of total Cx43 is restricted to the intercalated disk region of normal ventricular tissue. In ischemic heart, significant relocalization of total Cx43 to the lateral edges of myocytes was evident; however pS325/328/330-Cx43 remained predominately at the intercalated disk. Western blots indicated a eightfold decrease in pS325/328/330-Cx43 in ischemic tissue. Peptide-binding- and competition-experiments indicated that our antibody mainly detected Cx43 phosphorylated at S328 and/or S330 in heart tissue. To evaluate how this change in Cx43 phosphorylation contributes to ischemia-induced downregulation of intercellular communication, we stably transfected Cx43^-/- cells with a Cx43 construct in which serine residues 325, 328 and 330 had been mutated to alanine (Cx43-TM). Cx43-TM was not efficiently processed to isoforms that have been correlated with gap junction assembly. Nevertheless, Cx43-TM cells were electrically coupled, although development of coupling was delayed. Fully opened channels were only rarely observed in Cx43-TM cells, and Lucifer-Yellow-dye-coupling was significantly reduced compared with wild-type cells. These data suggest that phosphorylation of Cx43 at serine residues 325, 328 and/or 330 influences channel permselectivity and regulates the efficiency of gap junction assembly.

Key words: Connexin43, Gap junction, Heart, Ischemia, Phosphorylation

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

This article has been cited by other articles:

				M. Sato, Q. Jiao, T. Honda, R. Kurotani, E. Toyota, S. Okumura, T. Takeya, S. Minamisawa, S. M. Lanier, and Y. Ishikawa Activator of G Protein Signaling 8 (AGS8) Is Required for Hypoxia-induced Apoptosis of Cardiomyocytes: ROLE OF G{beta}{gamma} AND CONNEXIN 43 (CX43) J. Biol. Chem., November 6, 2009; 284(45): 31431 - 31440. [Abstract] [Full Text] [PDF]

				V. Akoyev, S. Das, S. Jena, L. Grauer, and D. J. Takemoto Hypoxia-Regulated Activity of PKC{epsilon} in the Lens Invest. Ophthalmol. Vis. Sci., March 1, 2009; 50(3): 1271 - 1282. [Abstract] [Full Text] [PDF]

				J. Qu, F. M. Volpicelli, L. I. Garcia, N. Sandeep, J. Zhang, L. Marquez-Rosado, P. D. Lampe, and G. I. Fishman Gap Junction Remodeling and Spironolactone-Dependent Reverse Remodeling in the Hypertrophied Heart Circ. Res., February 13, 2009; 104(3): 365 - 371. [Abstract] [Full Text] [PDF]

				R. P. Norris, M. Freudzon, L. M. Mehlmann, A. E. Cowan, A. M. Simon, D. L. Paul, P. D. Lampe, and L. A. Jaffe Luteinizing hormone causes MAP kinase-dependent phosphorylation and closure of connexin 43 gap junctions in mouse ovarian follicles: one of two paths to meiotic resumption Development, October 1, 2008; 135(19): 3229 - 3238. [Abstract] [Full Text] [PDF]

				N. J. Severs, A. F. Bruce, E. Dupont, and S. Rothery Remodelling of gap junctions and connexin expression in diseased myocardium Cardiovasc Res, October 1, 2008; 80(1): 9 - 19. [Abstract] [Full Text] [PDF]

				N. Kalcheva, J. Qu, N. Sandeep, L. Garcia, J. Zhang, Z. Wang, P. D. Lampe, S. O. Suadicani, D. C. Spray, and G. I. Fishman Gap junction remodeling and cardiac arrhythmogenesis in a murine model of oculodentodigital dysplasia PNAS, December 18, 2007; 104(51): 20512 - 20516. [Abstract] [Full Text] [PDF]

				J. L. Solan, L. Marquez-Rosado, P. L. Sorgen, P. J. Thornton, P. R. Gafken, and P. D. Lampe Phosphorylation at S365 is a gatekeeper event that changes the structure of Cx43 and prevents down-regulation by PKC J. Cell Biol., December 17, 2007; 179(6): 1301 - 1309. [Abstract] [Full Text] [PDF]