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PMID: 7841795 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Expression of recombinant rabbit UDP-GlcNAc: alpha 3-D-mannoside beta-1,2-N-acetylglucosaminyltransferase I catalytic domain in Sf9 insect cells.

Glycoconjugate journal ·Vol. 11 ·No. 3 ·1994-06-00 ·Pages 204-9

Sarkar M

Abstract

UDP-GlcNAc: alpha 3-D-mannoside beta-1,2-N-acetylglucosaminyltransferase I (GnT I; EC 2.4.1.101) catalyses a key reaction in the conversion of oligomannose to complex and hybrid N-glycans. The cytoplasmic tail and transmembrane segment of rabbit GnT I cDNA were replaced with an in-frame cleavable signal sequence and the hybrid construct was inserted into the genome of Autographa californica nuclear polyhedrosis virus (AcMNPV) under the control of the polyhedrin promoter. Sf9 insect cells were infected with the recombinant baculovirus and the enzymatically active and soluble catalytic domain of GnT I was purified from the medium (1-5 mg l-1) in two steps to a specific activity of about 2 mumol min-1 mg-1 protein. Recombinant GnT I has been used for the chemical-enzymatic synthesis of analogues of Man alpha 1-6]GlcNAc beta 1-2Man alpha 1-3]Man beta-O-octyl.

MeSH Terms
Animals Binding Sites Blotting, Southern Carbohydrate Sequence Catalysis Cells, Cultured Molecular Sequence Data N-Acetylglucosaminyltransferases/genetics,isolation & purification,metabolism Nucleopolyhedroviruses/genetics Rabbits Recombinant Proteins/genetics,isolation & purification,metabolism Spodoptera
Chemicals
Recombinant Proteins N-Acetylglucosaminyltransferases alpha-1,3-mannosyl-glycoprotein beta-1,2-N-acetylglucosaminyltransferase I
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Sarkar M
Research Institute, Hospital for Sick Children, Toronto, Ontario, Canada.
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Article Info
Journal
Glycoconjugate journal
Abbr.
Glycoconj J
ISSN
0282-0080
DOI
10.1007/BF00731219
Published
1994-06-00
Pages
204-9
Language
English
Region
United States
NLM ID
8603310
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00731219 View journal details
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