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Purification Protocol of β-Mannosyltransferase from Pig Aorta


Affiliations
1 Zoology Department, Meerut College, Meerut (UP), India
 

β-Mannosyltransferase that transfers mannose to Dol-PP-GlcNAc2 (chitobiose lipid acceptor) from GDP-Man was purified from the microsomes of pig aorta. The purification protocol consisted of the following steps: (i) preparation of microsomal fraction (ii) selective solubilization by Nonident NP-40 nonionic detergent at a protein to detergent ratio of 3:2; (iii) anion exchange chromatography on DE-52 in the presence of 0.4% NP-40, (iv)hydroxylapatite column chromatography, (v) chromatography on HQ anion exchange column, (vi) octyl Sepharose chromatography.

Keywords

β-Mannosyltransferase, Mannose, Dol-PP-GlcNAc2, GDP-Man, Chromatography, Pig Aorta, Protein-Detergent Ratio, Glycosyltransferase.
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  • Purification Protocol of β-Mannosyltransferase from Pig Aorta

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Authors

Seema Sharma
Zoology Department, Meerut College, Meerut (UP), India

Abstract


β-Mannosyltransferase that transfers mannose to Dol-PP-GlcNAc2 (chitobiose lipid acceptor) from GDP-Man was purified from the microsomes of pig aorta. The purification protocol consisted of the following steps: (i) preparation of microsomal fraction (ii) selective solubilization by Nonident NP-40 nonionic detergent at a protein to detergent ratio of 3:2; (iii) anion exchange chromatography on DE-52 in the presence of 0.4% NP-40, (iv)hydroxylapatite column chromatography, (v) chromatography on HQ anion exchange column, (vi) octyl Sepharose chromatography.

Keywords


β-Mannosyltransferase, Mannose, Dol-PP-GlcNAc2, GDP-Man, Chromatography, Pig Aorta, Protein-Detergent Ratio, Glycosyltransferase.