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J. Biol. Chem., Vol. 280, Issue 49, 40939-40947, December 9, 2005

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Tropoelastin Interacts with Cell-surface Glycosaminoglycans via Its COOH-terminal Domain^*

Thomas J. Broekelmann, Beth A. Kozel, Hideaki Ishibashi, Claudio C. Werneck, Fred W. Keeley, Lijuan Zhang¶, and Robert P. Mecham1

From the Departments of Cell Biology and Physiology and ^¶Pathology, Washington University School of Medicine, St. Louis, Missouri 63110 and the Cardiovascular Research Program, Research Institute, Hospital for Sick Children, Toronto, Ontario M5G 1X8, Canada

Using a biochemical and cell biological approach, we have identified a cell interaction site at the carboxyl terminus of tropoelastin. Cell interactions with the COOH-terminal sequence are not through the elastin-binding protein (EBP67) because neither VGVAPG-like peptides nor galactoside sugars altered adhesion. Our results also show that cell adhesion to tropoelastin is not promoted by integrins. Through the use of mutant Chinese hamster ovary cell lines defective in glycosaminoglycan biosynthesis, as well as competition studies and enzymatic removal of specific cell-surface glycosaminoglycans, the tropoelastin-binding moieties on the cell surface were identified as heparan and chondroitin sulfate-containing glycosaminoglycans, with heparan sulfate being greatly preferred. Heparin affinity chromatography combined with cell adhesion assays identified the last 17 amino acids as the sequence element at the carboxyl terminus of tropoelastin responsible for the adhesive activity.

Received for publication, July 6, 2005 , and in revised form, August 30, 2005.

^* This work was support by National Institutes of Health Grants HL53325, HL62295, and HL61006 (to R. P. M.), Pediatric Cardiology Training Grant T32 HL07873 (to B. A. K.), and the Heart and Stroke Foundation of Ontario (to F. W. K.). 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.

¹ To whom correspondence should be addressed: Campus Box 8228, 660 South Euclid Ave., St. Louis, MO 63110. Tel.: 314-362-2254; Fax: 314-362-2252; E-mail: bmecham{at}cellbiology.wustl.edu.

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