Solubilization of an α‐(1→3)‐D‐mannosyltransferase from pancreas which utilizes synthetic dolichyl pyrophosphate trisaccharide β‐Man‐(1→4)‐β‐GlcNAc‐(1→4)GlcNAc as substrate

FEBS Letters

Warren, Christopher D.¹  Jeanloz, Roger W.¹  Herscovics, Annette² 

[1] Laboratory for Carbohydrate Research, Department of Biological Chemistry and Medicine, Harvard Medical School and Massachusetts General Hospital, Boston, MA 02114, USA;McGill Cancer Centre, McGill University, Montreal, PQ H3G 1Y6, Canada

关键词: (1→3)-D-Mannosyltransferase Dolichyl pyrophosphate Tetrasaccharide biosynthesis;    Dol-PP-trisaccharide;    P1 -dolichyl P 2 -[O- β-D-mannopyranosyl-(1→4)-O-(2-acetamido-2-deoxy- β-D glucopyranosyl)(1→4)-2-acetamido-2-deoxy-β-D- glucopyranosyl]pyrophosphate;    Dol-PP-tetrasaccharide;    P1 -dolichyl-P 2 -[O-α-D-mannopyranosyl-(1→3)-O-β- D-mannopyranosyl-(1→4)-O-(2-acetamido-2-deoxy-β- D-glucopyranosyl-(1→4)-2-acetamido-2-deoxy-β-D- glucopyranosyl] pyrophosphate;    Dol-P-Man;    dolichyl β-D-mannopyranosyl phosphate;    Dol-P;    dolichyl phosphate;    BSA;    bovince serum albumin;   

DOI  :  10.1016/0014-5793(83)80516-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Calf pancreas microsomes were treated with 0.5–1% Triton X-100 and the resulting soluble enzyme preparation was incubated with GDP-D-[¹⁴C]mannose. The addition of synthetic Dol-PP derivative of the trisaccharide β-Man-(1→4)-β-GlcNac-(1→4)GlcNAc stimulated the synthesis of labeled lipid-bound tetrasaccharide 50–100-fold. The labeled tetrasaccharide thus formed was identified as α-Man-(1→3)-β-Man-(1→4)-β-GlcNAc-(1→4)GlcNAc by its chromatographic properties and by its sensitivity to α-mannosidase and to endo-β-N-acetylglucosaminidase D. The solubilized α-(1→3)mannosyltransferase did not require divalent cation and was active in the presence of 10 mM EDTA.

【 授权许可】

Unknown