Characterization of cell-associated proteoglycans synthesized by embryonic neural retinal cells.
To begin to understand the properties of the neuronal heparan sulfate proteoglycan (HSPG) that interacts with the neural cell adhesion molecule (NCAM), we have analyzed proteoglycans synthesized by dissociated embryonic retinal cells in culture. Because NCAM plays an integral role in cell-cell interactions, we have focused on cell-associated HSPGs. Proteoglycans were isolated from embryonic Day 10 retinal cell cultures labeled with 35SO4 and separated into fractions that either flowed through or were retained on phenyl-Sepharose. Molecules binding phenyl-Sepharose have been proposed to be capable of insertion into the plasma membrane, and thus may be involved in binding to NCAM. Proteoglycans binding to phenyl-Sepharose had an estimated molecular mass of 400-500 kDa, and contained 60% HSPG and 40% chondroitin sulfate proteoglycan (CSPG). The putative membrane-associated HSPGs, with an average molecular weight of 360 kDa, were shown to contain heparan sulfate chains of 40 and 20 kDa, and multiple core proteins with the major core protein having a molecular weight of approximately 130 kDa. The membrane-associated CSPGs also exhibited multiple core proteins, with sizes ranging from 120 to 220 kDa. These data suggest that multiple membrane-associated HSPGs and CSPGs are synthesized by embryonic neural retina cells, which may explain the diversity in function of this class of proteoglycans.
Burg
M
M
Department of Anatomy and Cell Biology, Medical University of South Carolina, Charleston 29425.
Cole
G J
GJ
D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D006025:Glycosaminoglycans / Q000096:biosynthesis / Q000302:isolation & purification; D007700:Kinetics; D008970:Molecular Weight; D009474:Neurons / Q000378:metabolism*; D011509:Proteoglycans / Q000096:biosynthesis* / Q000302:isolation & purification; D012160:Retina / Q000378:metabolism*
D006025:Glycosaminoglycans; D011509:Proteoglycans