Characterization of the oligosaccharide side chains on kainate binding proteins and AMPA receptors.
The amino acid sequences of the kainate binding proteins (KBPs) from frog and chicken brain are homologous with the carboxy terminal half of the rat brain AMPA receptors. In this study, we have characterized the oligosaccharide side chains present on the KBPs from chicken and frog brain, and the AMPA receptors (GluR1, GluR2, and GluR3) from rat brain. Deglycosylation of the asparagine-linked carbohydrates present on the chicken, frog, and rat receptor subunits with N-glycanase, resulted in decreases in the relative molecular weights (M(r)) of 3.4, 3.4, and 5.1 kDa respectively. Thus the percent of asparagine linked carbohydrate (based on M(r) values derived from SDS polyacrylamide gels) of the 49 kDa chicken, the 48 kDa frog, and the 107 kDa receptor rat subunits is 6.9, 7.1, and 4.8 percent respectively. No shifts in the M(r) were detected after treatment with neuraminidase indicating that sialic acid does not appear to be a major component of these receptors. Lectin binding studies demonstrated that both asparagine-linked and serine/threonine-linked oligosaccharides were present in the chicken, frog, and rat proteins. The data indicate that at least one of the asparagine linked oligosaccharide side chains appear to be of the complex or non-bisected hybrid type in all three species. The similarities in the glycosyl moieties of the chicken and frog kainate KBPs and the rat brain AMPA receptors suggests that the homology in the amino acid sequences between these proteins may extend to homology in their oligosaccharide sides chains as well.
Hullebroeck
M F
MF
Faculty of Pharmacy, University of Toronto, Ont, Canada.
Hampson
D R
DR
D000581:Amidohydrolases; D000818:Animals; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D007608:Kainic Acid; D037102:Lectins; D009439:Neuraminidase; D009844:Oligosaccharides / Q000032:analysis*; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D011894:Rana pipiens; D051381:Rats; D017208:Rats, Wistar; D018091:Receptors, AMPA; D018092:Kainic Acid Receptors; D017981:Receptors, Neurotransmitter / Q000737:chemistry* / Q000302:isolation & purification; D017385:Sequence Homology; D013045:Species Specificity
D000581:Amidohydrolases; D007608:Kainic Acid; D037102:Lectins; D009439:Neuraminidase; D009844:Oligosaccharides; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D018091:Receptors, AMPA; D018092:Kainic Acid Receptors; D017981:Receptors, Neurotransmitter