Ž . Matrix Biology 18 1999 163178 Human perlecan immunopurified from different endothelial cell sources has different adhesive properties for vascular cells John M. Whitelock a,b,  , Lloyd D. Graham b , James Melrose c , Alan D. Murdoch d,1 , Renato V. Iozzo d , P. Anne Underwood a,b a ( ) Co-operati  e Research Center CRC for Cardiac Technology, North Ryde, NSW 1670, Australia b CSIRO Molecular Science, Sydney Laboratory, P.O. Box 184, North Ryde, NSW 1670, Australia c Raymond Pur  es Research Laboratories, Royal North Shore Hospital, St Leonards, NSW 2065, Australia d Dept. Pathology, Anatomy and Cell Biology, and The Kimmel Cancer Center, Thomas Jefferson Uni  ersity, Philadelphia PA 19107 USA Accepted 4 January 1999 Abstract Perlecan, a major heparan sulfate proteoglycan of vascularized tissues, was immunopurified from media conditioned by human endothelial cells of both arterial and venous origin. The heparan sulfate moiety of perlecan from cultured arterial cells differed in amount andor composition from that produced by a transformed cell line of venous origin. Both forms of perlecan bound basic fibroblast growth factor with K 70 nM. In ELISA experiments, perlecan and its protein core bound to d various extracellular matrix components in a manner that was strongly influenced by the format of the assay. Human vascular smooth muscle cells and human endothelial cells adhered to perlecan-coated surfaces, and both cell types adhered better to the venous cell-derived than to the arterial cell-derived perlecan. Removal of the heparan sulfate chains abolished this difference and increased the ability of both types of perlecan to adhere vascular cells. Denaturation of perlecan and its protein core also rendered each of them more adhesive, indicating the presence of conformation-independent adhesion determinants in the polypeptide sequence. Their location was investigated using recombinant perlecan domains. Overall, our results represent the first demonstration of human perlecan acting as an adhesive molecule for human vascular cells and suggest that it may play a role in vascular wound healing.  1999 Elsevier Science B.V.International Society of Matrix Biology. All rights reserved. Keywords: Adhesion; Endothelial; Human; Perlecan; Smooth muscle 1. Introduction Ž . Perlecan is the major heparan sulfate HS proteog- lycan of basement membranes and other connective  Corresponding author. CSIRO Molecular Science, Sydney Laboratory, P.O. Box 184, North Ryde, Sydney, NSW 2113, Aus- tralia. Tel.: 61 2 94905055; fax: 61 2 94905002; e-mail: john.whitelock@molsci.csiro.au 1 Current address: Wellcome Trust Centre for Cell Matrix Re- search, School of Biological Sciences, University of Manchester, Manchester, M13 9PT, UK. tissues. Perlecan has been purified from the mouse Ž . Ž Englebroth Holm Swarm EHS tumor Hassell et al., 1980; Paulsson et al., 1987; Saku and Furthmayr, 1989; Hayashi et al., 1992; Battaglia et al., 1993; Klein . Ž . et al., 1995 , porcine kidneys Gauer et al., 1996 , Ž . bovine kidneys Hagen et al., 1993 , human placenta Ž . Ž Isemura et al., 1987 , human carcinoma cells Iozzo, . 1984 , from the conditioned medium of bovine aortic Ž . endothelial cells Saku and Furthmayr, 1989 , and from the extracellular matrix of cultured human fetal Ž lung fibroblasts Heremans et al., 1988; Aviezer et al., . 1994 . In all cases, the perlecan was shown to be a 0945-053X99$ - see front matter  1999 Elsevier Science B.V.International Society of Matrix Biology. All rights reserved. Ž . PII: S 0 9 4 5 - 0 5 3 X 99 00014-1