Characterization of gp93, a novel, highly heterogeneous glycoprotein present in growth cone membranes.
gp93 was first described in growth cones from fetal rat brain as a 90-97-kDa glycoprotein family that binds wheat-germ agglutinin and consists of at least 12 different isoelectric variants (pl range approximately 4.9-6.4). Of particular interest is that different sets of gp93 variants are expressed in growth cones isolated from different brain regions. The preparation of a polyclonal antibody to gp93 allowed further characterization of this glycoprotein. The carbohydrate groups of gp93 were partially characterized by digestion with different glycosidases. The results indicate that most or all oligosaccharide units are N-linked (asparagine-linked) and contain sialic acid. Two-dimensional polyacrylamide gel electrophoresis and western blot with anti-gp93 show that deglycosylated gp93 is an only slightly heterogeneous polypeptide of 66 kDa, indicating that gp93 heterogeneity is due, primarily or exclusively, to differential glycosylation. Analysis of the tissue distribution in fetal rat showed gp93 to be highly enriched in the brain. Immunoblots and immunostaining of cross sections of developing cerebellum revealed that gp93 is developmentally regulated in this tissue, associated primarily with growing parallel fibers and Purkinje dendrites. Immunostaining of neurons in culture shows significant amounts of gp93 in elongating neurites and growth cones. Our results indicate that gp93 is a developmentally regulated glycoprotein of the brain that is most prominent in growth cones and growing neurites and that appears to be glycosylated differentially by different neurons.
Quiroga
S
S
Department of Cellular and Structural Biology, University of Colorado School of Medicine, Denver.
Pfenninger
K H
KH
D000818:Animals; D000918:Antibody Specificity; D001921:Brain / Q000196:embryology*; D001923:Brain Chemistry; D002462:Cell Membrane / Q000737:chemistry; D002478:Cells, Cultured; D002531:Cerebellum / Q000737:chemistry / Q000254:growth & development; D003712:Dendrites / Q000737:chemistry; D005455:Fluorescent Antibody Technique; D006031:Glycosylation; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D009412:Nerve Fibers / Q000737:chemistry; D009474:Neurons / Q000737:chemistry; D011689:Purkinje Cells / Q000737:chemistry; D051381:Rats; D014018:Tissue Distribution
D008562:Membrane Glycoproteins