Intramolecular heterogeneity of degradation in plasma membrane glycoproteins: evidence for a general characteristic.
Five integral plasma membrane glycoproteins (60, 80, 120, 140, and 160 kilodaltons) were isolated to homogeneity from rat liver by a four-step procedure: (i) extraction of plasma membranes with lithium diiodosalicylate, (ii) solubilization of glycoproteins with Nonidet P-40, (iii) affinity chromatography on concanavalin A-Sepharose, and (iv) semipreparative NaDodSO4/polyacrylamide gel electrophoresis. The glycoproteins contained 48.5--51.5% hydrophobic amino acids. Carbohydrate moieties contained N-acetyl-D-glucosamine, D-mannose, D-galactose, L-fucose, and N-acetylneuraminic acid. N-Acetyl-D-galactosamine was not detectable. Half-lives of degradation of the carbohydrate and protein moieties of the five glycoproteins were measured by pulse-chase experiments in vivo. Protein moieties had half-lives ranging from 52 to 88 hr in the five glycoproteins, with a mean of 73 +/- 15 hr. Terminal sugars, L-fucose, and N-acetylneuraminic acid had significantly shorter half-lives, averaging 18 +/- 2 hr and 29 +/- 3 hr, respectively. The half-life of D-mannose varied between that of the terminal sugars and that of the protein moiety, depending on the type of the glycoprotein. The data show that the carbohydrate moieties are degraded faster than the protein portion of the glycoproteins. As this finding was obtained in each of the five glycoproteins, intramolecular heterogeneity of breakdown may be a general characteristic of plasma membrane glycoproteins in liver.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002462:Cell Membrane / Q000378:metabolism*; D006023:Glycoproteins / Q000302:isolation & purification / Q000378:metabolism*; D006207:Half-Life; D007700:Kinetics; D008099:Liver / Q000378:metabolism*; D008297:Male; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
D000596:Amino Acids; D002241:Carbohydrates; D006023:Glycoproteins; D008565:Membrane Proteins