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

J. Biol. Chem., Vol. 280, Issue 23, 22425-22436, June 10, 2005

This Article Full Text Full Text (PDF) All Versions of this Article: 280/23/22425    most recent M413380200v1 Alert me when this article is cited 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 Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Chen, D. Articles by Langford, M. P. Search for Related Content PubMed PubMed Citation Articles by Chen, D. Articles by Langford, M. P. Social Bookmarking                      What's this?

Surface Calreticulin Mediates Muramyl Dipeptide-induced Apoptosis in RK₁₃ Cells^*

Dequan Chen, Donald E. Texada, Chris Duggan, Chanping Liang, Thomas B. Reden, Lakshmana M. Kooragayala, and Marlyn P. Langford¶

From the Departments of Ophthalmology and ^¶Microbiology & Immunology, Louisiana State University Health Sciences Center, Shreveport, Louisiana 71130

Calreticulin (CRT) is a binding protein for apoptotic N-acetylmuramyl-L-alanyl-D-isoglutamine (L,D-MDP) or peptidoglycan in RK₁₃ cells. CRT on RK₁₃ cell surface (srCRT) forms complex(es) with tumor necrosis factor receptor 1 (TNFR1) and TNFR-associated death domain (TRADD) protein of the cell membrane. CRT polyclonal or monoclonal antibody binding to RK₁₃ srCRT dose-dependently inhibited L,D-MDP-induced apoptosis. In RK₁₃ cells, L,D-MDP up-regulated the TNFR1·TRADD complex of the plasma membrane and subsequently induced cytosolic TRADD-Fas-associated death domain protein complex. Biotinylated srCRT was capable of calcium-dependent binding of Sepharose-immobilized L,D-MDP or peptidoglycan. However, Toll-like receptors TLR-2 and TLR-4, Nod2, and CD14 of RK₁₃ cells did not specifically bind Sepharose-immobilized L,D-MDP. High concentrations (5-40 mM) of EGTA dose-dependently inhibited free L,D-MDP binding to purified RK₁₃ cell CRT and promoted free L,D-MDP dissociation from RK₁₃ cell CRT·MDP complex. Different concentrations of EGTA (0-40 mM) added to Dulbecco's modified essential medium with 1.8 mM calcium or phosphate-buffered saline with 0.18 mM calcium have different effects on medium free calcium concentrations but have identical inhibiting effects on L,D-MDP-induced apoptosis. More inhibition of the L,D-MDP-induced apoptotic DNA ladders and caspase-3 activity in RK₁₃ cells was obtained with EGTA pretreatment (83%) than just EGTA + L,D-MDP (47%). The knocking down of srCRT by antisense oligonucleotide CRTAS121 (250 nmol/ml) and stealth small interfering RNA CRT_siR479 (150 pM/ml) for 2 days (44 and 66%, respectively), resulted in the inhibition of L,D-MDP-induced caspase-3 activity (47 and 65%, respectively). The results suggest that (a) the binding of L,D-MDP to srCRT is calcium-dependent, i.e. on srCRT-bound calcium, and (b) it is srCRT, not TLR-2, TLR-4, Nod2 or CD14, that mediates L,D-MDP-induced RK₁₃ cell apoptosis through activating the TNFR1· TRADD-Fas-associated death domain protein apoptotic pathway.

Received for publication, November 29, 2004 , and in revised form, March 24, 2005.

^* This work was supported by the Eye Clinic and Dept. of Ophthalmology (Louisiana Health Sciences Center, Shreveport, LA). 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.

To whom correspondence should be addressed: Dept. of Ophthalmology, Louisiana State University Health Sciences Center, 1501 Kingshighway, Shreveport, LA 71130-3932. Tel.: 318-675-5010; Fax: 318-675-6000; E-mail: dchen{at}lsuhsc.edu.

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

This article has been cited by other articles:

				X.-Y. Jia, C.-Y. Xu, R.-L. Jing, R.-Z. Li, X.-G. Mao, J.-P. Wang, and X.-P. Chang Molecular cloning and characterization of wheat calreticulin (CRT) gene involved in drought-stressed responses J. Exp. Bot., March 1, 2008; 59(4): 739 - 751. [Abstract] [Full Text] [PDF]

				M. P. Langford, T. B. Redens, N. R. Harris, S. Lee, S. K. Jain, S. Reddy, and R. McVie Plasma Levels of Cell-Free Apoptotic DNA Ladders and Gamma-Glutamyltranspeptidase (GGT) in Diabetic Children Experimental Biology and Medicine, October 1, 2007; 232(9): 1160 - 1169. [Abstract] [Full Text] [PDF]

				G. Zeng, M. E. Aldridge, X. Tian, D. Seiler, X. Zhang, Y. Jin, J. Rao, W. Li, D. Chen, M. P. Langford, et al. Dendritic Cell Surface Calreticulin Is a Receptor for NY-ESO-1: Direct Interactions between Tumor-Associated Antigen and the Innate Immune System J. Immunol., September 15, 2006; 177(6): 3582 - 3589. [Abstract] [Full Text] [PDF]

				S. Traub, S. von Aulock, T. Hartung, and C. Hermann Invited review: MDP and other muropeptides direct and synergistic effects on the immune system Innate Immunity, April 1, 2006; 12(2): 69 - 85. [Abstract] [PDF]

				D. Weichart, J. Gobom, S. Klopfleisch, R. Hasler, N. Gustavsson, S. Billmann, H. Lehrach, D. Seegert, S. Schreiber, and P. Rosenstiel Analysis of NOD2-mediated Proteome Response to Muramyl Dipeptide in HEK293 Cells J. Biol. Chem., January 27, 2006; 281(4): 2380 - 2389. [Abstract] [Full Text] [PDF]