Purification of a 58-kDa protein (ER58) from monkey liver microsomes and comparison with protein-disulfide isomerase.
A 58-kDa protein (ER58) was purified from monkey liver to apparent homogeneity. It accounts for more than 3% of microsomal proteins and is highly conserved among several mammalian species. The amino acid compositions of the N-terminal part and that of two internal peptide fragments present strong similarities with the sequence ascribed to phospholipase C-alpha. Numerous proteins exhibiting a high similarity with this sequence have been isolated by other investigators. Their biological function is controversial. Our purified protein is not active as a phosphatidylinositol-specific phospholipase C, protease or carnitine acyl transferase. Although less efficient than authentic protein-disulfide isomerase, ER58 catalyses the glutathione-dependent reduction of insulin and the reorganization of disulfide bonds of randomly oxidized (scrambled) ribonuclease in reducing conditions. In contrast, ER58 is devoid of oxidizing activity on thiol groups of reduced proteins. Many studies suggest that the proteins bearing the phospholipase C-alpha sequence could be considered as protein-disulfide isomerase isozymes. Our results indicate that ER58 is not totally similar to protein-disulfide isomerase in performing thiol :protein-disulfide oxidoreductase reactions and suggest that the two proteins may exert distinct cellular functions.
Bonfils
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C
INSERM, Unité 128, CNRS, Montpellier, France. bonfils@xerxes.crbm.cnrs-mop.fr
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D066298:In Vitro Techniques; D008862:Microsomes, Liver / Q000737:chemistry* / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009113:Muramidase; D010215:Papio; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D054801:Phosphoinositide Phospholipase C; D019704:Protein Disulfide-Isomerases / Q000378:metabolism*; D011506:Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D012260:Ribonucleases; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D010738:Type C Phospholipases / Q000235:genetics / Q000378:metabolism
D011506:Proteins; D012260:Ribonucleases; D010738:Type C Phospholipases; D054801:Phosphoinositide Phospholipase C; D009113:Muramidase; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D019704:Protein Disulfide-Isomerases