Y2 receptor proteins for peptide YY and neuropeptide Y. Characterization as N-linked complex glycoproteins.
Affinity labeling using [125I-Tyr36]PYY and homobifunctional affinity crosslinking reagents of the rabbit Y2 receptor for peptide YY(PYY) results in specifically labeled proteins of both M(r) = 50,000 to 60,000 and M(r) = 96,000 to 115,000 [1,2]. In this work the glycoprotein nature of affinity labeled Y2 receptor proteins were investigated by enzymatic deglycosylation using neuraminidase, endoglycosidase F (endo F), N-glycosidase F (PNGase F), and O-glycanase treatment. Only N-glycosidase F and neuraminidase increased the electrophoretic mobility of the radiolabeled receptor bands, whereas all other glycosidases did not. PNGase F treatment of both radiolabeled receptor bands electroeluted from gel slices reduced the apparent molecular mass of by 16-17 kDa units, that is M(r) = 96,000 to 79,000 and M(r) = 60,000 to 44,000, indicating removal of N-linked oligosaccharide chains of similar size from both species. Neuraminidase treatment caused slight increases in the electrophoretic mobilities suggesting the presence of terminal sialic residues. It is concluded that the Y2 binding proteins are N-linked complex (sialo)glycoproteins with a minimal core protein size of M(r) = 44,000. Furthermore, based on this sensitivity pattern of the glycosidases, the Asn-linked carbohydrate may be of the tri- or tetra-antennary complex type containing terminal sialic acid residues.
Hansen
A P
AP
Department of Clinical Biochemistry, University of Copenhagen, Denmark.
Sheikh
S P
SP
D000345:Affinity Labels; D000818:Animals; D001216:Asparagine / Q000378:metabolism; D050260:Carbohydrate Metabolism; D005260:Female; D006023:Glycoproteins / Q000378:metabolism*; D007687:Kidney Tubules, Proximal / Q000378:metabolism; D009439:Neuraminidase / Q000378:metabolism; D009478:Neuropeptide Y / Q000378:metabolism*; D019894:Peptide YY; D010455:Peptides / Q000378:metabolism*; D011817:Rabbits; D017476:Receptors, Neuropeptide Y; D017981:Receptors, Neurotransmitter / Q000737:chemistry / Q000378:metabolism*
D000345:Affinity Labels; D006023:Glycoproteins; D009478:Neuropeptide Y; D010455:Peptides; D017476:Receptors, Neuropeptide Y; D017981:Receptors, Neurotransmitter; D019894:Peptide YY; D001216:Asparagine; D009439:Neuraminidase