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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
Author(s) -
Herscovics Annette,
Warren Christopher D.,
Jeanloz Roger W.
Publication year - 1983
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(83)80516-x
Subject(s) - trisaccharide , chemistry , tetrasaccharide , stereochemistry , mannose , oligosaccharide , substrate (aquarium) , biochemistry , polysaccharide , oceanography , geology
Calf pancreas microsomes were treated with 0.5–1% Triton X‐100 and the resulting soluble enzyme preparation was incubated with GDP‐D‐[ 14 C]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.