Aldosterone-induced glycoproteins: further characterization.
Aldosterone induces the synthesis of a group of glycoproteins (GP65,70) in toad urinary bladders which are potential effectors of the natriferic action of this hormone. The GP65,70 complex is composed of two molecular weight classes of proteins (Mr 65 and 70 kDa), each class being composed of several discrete proteins of varying isoelectric points (5.8-6.2). These proteins can be partially enriched (approximately 20-fold) using wheat germ agglutinin-sepharose affinity chromatography, are neuraminidase-resistant, and can be N-deglycosylated by endoglycosidase-H and N-glycanase. Treatment with N-glycanase leads to the appearance of a microheterogeneous group of proteins, all having the same Mr (approximately 40 kDa). From these studies it can be concluded that these particular aldosterone-induced proteins: (1) are heavily glycosylated, (2) contain multiple high mannose and hybrid oligosaccharides side chains, and (3) contain similar (if not identical) peptide backbones. Post-translational N-glycosylation accounts, at least in part, for their electrophoretic polymorphism (variation in Mr) but not for their electrophoretic microheterogeneity (variation in pI). The latter may reflect other types of post-translational modification (e.g. O-glycosylation, phosphorylation) or may be due to subtle differences in amino acid composition. The partial purification and biochemical characterization of GP65,70 should ultimately lead to a better understanding of the function of these putative "effectors" of aldosterone-stimulated Na+ transport.
Szerlip
H M
HM
Medical Service, Philadelphia Veterans Administration Medical Center, PA 19104.
Cox
M
M
D000450:Aldosterone / Q000494:pharmacology*; D000818:Animals; D002024:Bufo marinus; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006023:Glycoproteins / Q000096:biosynthesis*; D006026:Glycoside Hydrolases; D006596:Hexosaminidases; D009439:Neuraminidase; D001743:Urinary Bladder / Q000187:drug effects* / Q000378:metabolism
D006023:Glycoproteins; D000450:Aldosterone; D006026:Glycoside Hydrolases; D006596:Hexosaminidases; C020668:glycanase; D009439:Neuraminidase