Conserved structural features in glycoprotein processing glucosidase I from several tissues and species.
Glucosidase I initiates the processing of the oligosaccharide, Glc3Man9GlcNAc2, in newly assembled glycoproteins by excising the distal alpha 1,2-linked glucosyl residue in the oligosaccharide. Earlier, the enzyme purified from the ER of rat and bovine mammary gland has been found to have M(r) of 85 kDa, as examined by SDS-PAGE along with a domain structure in which a 39 kDa lumenally-oriented region is anchored to the ER through a transmembrane segment and a short cytoplasmic tail. These studies were further extended to include the enzyme from several different tissues of the rat, mouse, guinea pig and bovine mammary glands, sheep liver and pig kidney. Using anti-rat glucosidase I antibody as a probe and several biochemical parameters such as SDS-PAGE analysis, trypsin-catalyzed digestion, ConA-binding, endo H susceptibility and peptide mapping analysis by cleavage of the tryptophanyl peptide linkages within the enzyme, it was found that glucosidase I in all of the tissue sources examined has an M(r) of 85 kDa and is cross-reactive to anti-rat glucosidase antibody. The enzyme is a high mannose glycoprotein, and has domain features in its structure; the enzyme from mouse, rat, guinea pig and bovine mammary glands and sheep liver is sequentially cleaved by trypsin to generate fragments of 69, 55 and 39 kDa. The rate of release of the different fragments differs for different sources, indicating some evolutionary changes in its primary structure. The trypsin-released fragments from pig kidney enzyme are 69, 45 and 29 kDa in size, identical to the same observed earlier for pig liver.(ABSTRACT TRUNCATED AT 250 WORDS)
Pukazhenthi
B S
BS
Department of Animal Sciences, University of Maryland, College Park 20742.
Varma
G M
GM
Vijay
I K
IK
D000818:Animals; D002240:Carbohydrate Sequence; D002417:Cattle; D005260:Female; D006023:Glycoproteins / Q000096:biosynthesis*; D006168:Guinea Pigs; D008099:Liver / Q000201:enzymology; D008321:Mammary Glands, Animal / Q000201:enzymology; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009844:Oligosaccharides / Q000378:metabolism*; D009928:Organ Specificity; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D011499:Protein Processing, Post-Translational; D051381:Rats; D012756:Sheep; D013045:Species Specificity; D013552:Swine; D000520:alpha-Glucosidases / Q000737:chemistry* / Q000378:metabolism
D006023:Glycoproteins; D009844:Oligosaccharides; D010446:Peptide Fragments; C042059:glucosidase I; D000520:alpha-Glucosidases