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Isolation and partial characterization of a cell‐surface heparan sulfate proteoglycan from embryonic rat spinal cord
Author(s) -
Guiseppetti J. M.,
McCarthy J. B.,
Letourneau P. C.
Publication year - 1994
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.490370505
Subject(s) - heparan sulfate , proteoglycan , fibronectin , perlecan , chemistry , glycosaminoglycan , affinity chromatography , extracellular matrix , neurite , biochemistry , cell adhesion molecule , neuron , microbiology and biotechnology , neural cell adhesion molecule , cell adhesion , cell , biology , in vitro , enzyme , neuroscience
Cell‐surface heparan sulfate proteoglycans (HSPGs) are potential mediators of neuronal cell adhesion, spreading, and neurite outgrowth on various extracellular matrix molecules. One possible site of HSPG attachment is a heparin binding domain of fibronectin, which is present in the synthetic peptide FN‐C/H II. In this study, HSPGs extracted from embryonic rat spinal cord by detergent were purified by ionexchange chromatography, gel filtration, and affinity chromatography on an agarose column coupled with FN‐C/H II conjugated to ovalbumin (OA). Heparitinase treatment of the iodinated HSPG fraction led to the appearance of a major protein core with a molecular size of 72 kDa, as determined by reducing SDS‐PAGE. The intact proteoglycan has a molecular size of approximately 150–165 kDa, containing heparan sulfate glycosaminoglycan chains of about 10–15 kDa. Anti‐HSPG antibodies recognized the 72 kDa core protein by immunoblotting, and stained the surface of spinal cord neurons, oligodendrocytes, and a subset of astrocytes. These results identify a cell‐surface HSPG that may mediate neuron‐substratum or neuron‐glia interactions in embryonic central nervous system. © 1994 Wiley‐Liss, Inc.