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J Biol Chem, Vol. 273, Issue 19, 11483-11490, May 8, 1998

Cell Surface Glycosaminoglycans Do Not Serve as Ligands for PECAM-1
PECAM-1 IS NOT A HEPARIN-BINDING PROTEIN

Qi-Hong Sun, Cathy Paddock, Gian P. Visentin, Mark M. Zukowski^§, William A. Muller^¶, and Peter J. Newman^**

From the  Blood Research Institute, The Blood Center of Southeastern Wisconsin, Milwaukee, Wisconsin 53233-2194, ^§ Amgen Boulder, Inc., Boulder, Colorado 80301, the ^** Departments of Cellular Biology and Pharmacology, The Medical College of Wisconsin, Milwaukee, Wisconsin 53226, and the ^¶ Department of Pathology, Cornell University Medical College, New York, New York 10021

Previous studies have suggested that PECAM-1 mediates cellular interactions via both homophilic and heterophilic adhesive mechanisms. Cell surface glycoaminoglycans have been implicated as one of the heterophilic ligands for PECAM-1. To determine whether PECAM-1 is capable of interacting directly with glycosaminoglycans, we examined the adhesive properties of multiple monovalent and multivalent forms of this adhesion molecule. We found that the binding of a bivalent PECAM-1/IgG chimeric protein or multivalent PECAM-1-containing proteoliposomes to multiple different cell lines was 1) strictly dependent upon cell surface expression of PECAM-1 and 2) unaffected by the presence of excess heparin or heparan sulfate. The extracellular domain of PECAM-1 failed to interact specifically with heparin-Sepharose, ³H-labeled heparin, or a heparin-bovine serum albumin conjugate. In addition, an amino acid sequence motif inadvertently created by the juxtaposition of PECAM-1 and IgG sequences within the hinge region of certain PECAM-1/IgG chimeric constructs was found to confer glycosaminoglycan binding properties not normally present within the extracellular domain of the native molecule. Together, these data suggest that the mechanism by which heparin is able to affect PECAM-1-dependent cell-cell adhesion is indirect and occurs via inhibition of events that occur downstream from PECAM-1 engagement.

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Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.

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