Analysis of proteoglycan expression in developing chicken brain: characterization of a heparan sulfate proteoglycan that interacts with the neural cell adhesion molecule

Abstract

In the present study we have characterized the major proteoglycans of chick brain, focusing on their pattern of expression in development and on identifying the heparan sulfate proteoglycan (HSPG) that binds to the neural cell adhesion molecule (NCAM). The major chondroitin sulfate proteoglycans (CSPG) are a heterogeneous group of molecules with an average MW of 450 kDa. Protein core analysis reveals multiple protein cores between 100 and 350 kDa. The HSPGs are somewhat smaller, with an average MW of 350 kDa, and the major brain HSPG possesses a 250 kDa protein core. During development the relative percentage of HSPG decreases from approximately 50% of total sulfate-labeled PG at E6 to 25% by E10. In order to begin to characterize the HSPG that interacts with NCAM, we initially used an antiserum produced against a HSPG which was previously shown to copurify with NCAM (Cole and Burg: Exp Cell Res 182:44-60, 1989). This antiserum immunoprecipitated a HSPG core protein of 250 kDa, corresponding to the major HSPG of chick brain. We also show that the major brain HSPG binds to a synthetic peptide that encodes the heparan sulfate-binding domain of NCAM, and that monoclonal antibodies to a recently identified chick retinal HSPG recognize this NCAM-binding HSPG. This HSPG was immunopurified from E10 chick brain using the 6D2 monoclonal antibody, and has been shown to bind an affinity column containing the heparan sulfate-binding peptide of NCAM. Consistent with its ability to bind NCAM, we show that the intact 6D2 HSPG inhibits cell adhesion to a HBD peptide substratum, and also binds chick brain cells when employed as a substratum.