The Proteoglycan Lectin Domain Binds Sulfated Cell Surface Glycolipids and Promotes Cell Adhesion

doi:10.1074/jbc.274.16.11431

The Proteoglycan Lectin Domain Binds Sulfated Cell Surface Glycolipids and Promotes Cell Adhesion*

From the ^‡Burnham Institute, La Jolla, California 92037 and the ^¶Department of Pharmacology and Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205

Abstract

The lecticans are a group of chondroitin sulfate proteoglycans characterized by the presence of C-type lectin domains. Despite the suggestion that their lectin domains interact with carbohydrate ligands, the identity of such ligands has not been elucidated. We previously showed that brevican, a nervous system-specific lectican, binds the surface of B28 glial cells (Yamada, H., Fredette, B., Shitara, K., Hagihara, K., Miura, R., Ranscht, B., Stallcup, W. B., and Yamaguchi, Y. (1997) J. Neurosci. 17, 7784–7795). In this paper, we demonstrate that two classes of sulfated glycolipids, sulfatides and HNK-1-reactive sulfoglucuronylglycolipids (SGGLs), act as cell surface receptors for brevican. The lectin domain of brevican binds sulfatides and SGGLs in a calcium-dependent manner as expected of a C-type lectin domain. Intact, full-length brevican also binds both sulfatides and SGGLs. The lectin domain immobilized as a substrate supports adhesion of cells expressing SGGLs or sulfatides, which was inhibited by monoclonal antibodies against these glycolipids or by treatment of the substrate with SGGLs or sulfatides. Our findings demonstrate that the interaction between the lectin domains of lecticans and sulfated glycolipids comprises a novel cell substrate recognition system, and suggest that lecticans in extracellular matrices serve as substrate for adhesion and migration of cells expressing these glycolipids in vivo.

Footnotes

↵* This work was supported by National Institutes of Health Grants NS32717 and HD25938 (to Y. Y.) and CA28896 (to E. R.), and by Cancer Center Support Grant CA30199.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵§ Current address: Dept. of Cell and Molecular Biology, Lund University, P. O. Box 94, S-221 00 Lund, Sweden.
↵‖ To whom correspondence should be addressed: Burnham Institute, 10901 N. Torrey Pines Rd., La Jolla, CA 92037. Tel.: 619-646-3124; Fax: 619-646-3199; E-mail: yyamaguchi{at}burnham-inst.org.
↵2 M. Fukuda, personal communication.
Abbreviations:

CSPG

chondroitin sulfate proteoglycan

PLD

proteoglycan lectin domain

CRP

complement regulatory protein

ECM

extracellular matrix

SGGL

sulfoglucuronylglycolipid

rCLD

recombinant C-type lectin domain

FNIII

fibronectin type III

EGF

epidermal growth factor

CHO

Chinese hamster ovary

TBS

Tris-buffered saline

MDCK

Madin-Darby canine kidney

HRP

horseradish peroxidase

PBS

phosphate-buffered saline

BSA

bovine serum albumin

CMF

calcium- and magnesium-free

HBSS

Hank’s balanced salt solution
- Received December 24, 1998.
The American Society for Biochemistry and Molecular Biology, Inc.