PMID: 3322267Nov 1, 1987Paper

Purification and characterization of glucosidase I involved in N-linked glycoprotein processing in bovine mammary gland

The Biochemical Journal
K ShailubhaiI K Vijay

Abstract

Glucosidase I, the first enzyme involved in the post-translational processing of N-linked glycoproteins, was purified to homogeneity from the lactating bovine mammary tissue. The enzyme was extracted by differential treatment of the microsomal fraction with Triton X-100 and Lubrol PX. The solubilized enzyme was subjected to affinity chromatography on Affi-Gel 102 with N-5-carboxypentyldeoxynojirimycin as ligand and DEAE-Sepharose CL-6B chromatography. Purified glucosidase I shows a molecular mass of 320-330 kDa by gel filtration on Sephacryl S-300. SDS/polyacrylamide-gel electrophoresis under reducing conditions indicates a single band of approx. 85 kDa, indicating that the native enzyme is probably a tetrameric protein. Several criteria, including pH optimum of 6.6-7.0, specific hydrolytic action towards Glc3Man9GlcNAc2, to release the terminally alpha-1,2-linked glucosyl residue, and total lack of activity towards Glc1Man9GlcNAc2 and Glc2Man9GlcNAc2 saccharides, which are the biological substrates for processing glucosidase II, and 4-methylumbelliferyl alpha-D-glucopyranoside show the non-lysosomal origin and the processing-specific role of the purified enzyme. The enzyme does not require any metal ions for its activity. Hg2+...Continue Reading

Citations

May 2, 2000·American Journal of Human Genetics· De Praeter CM Van Coster RN
Aug 19, 2014·Journal of Neuroscience Research·Antje AnjiMeena Kumari
Jul 16, 1999·Glycobiology·F A KhanI K Vijay
Oct 1, 1993·Biological Chemistry Hoppe-Seyler·J M AlonsoP Calvo
Nov 24, 1998·Critical Reviews in Oral Biology and Medicine : an Official Publication of the American Association of Oral Biologists·M A Kukuruzinska, K Lennon
Aug 15, 1989·Analytical Biochemistry·M S KangP S Sunkara
May 1, 1990·Bio/technology·C F Goochee, T Monica
Jul 15, 1995·European Journal of Biochemistry·B Kalz-FüllerE Bause
Apr 2, 2010·Antonie van Leeuwenhoek·María D Frade-PérezHéctor M Mora-Montes
Apr 17, 2001·The Journal of Biological Chemistry·D Heritage, W F Wonderlin
Apr 24, 2018·Biochemistry·Keya MukherjeeFrank M Raushel

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