LEHLE, L and EIDEN, A and LEHNERT, K and HASELBECK, A and KOPETZKI, E (1995) GLYCOPROTEIN-BIOSYNTHESIS IN SACCHAROMYCES-CEREVISIAE - NGD29, AN N-GLYCOSYLATION MUTANT ALLELIC TO OCH1 HAVING A DEFECT IN THE INITIATION OF OUTER CHAIN FORMATION. FEBS LETTERS, 370 (1-2). pp. 41-45. ISSN 0014-5793,
Full text not available from this repository.Abstract
Outer drain glycosylation in Saccharomyces cerevisiae leads to heterogeneous and immunogenic asparagine-linked saccharide chains containing more than 50 mannose residues on secreted glycoproteins. Using a [H-3]mannose suicide selection procedure a collection of N-glycosylation defective mutants (designated ngd) was isolated. One mutant, ngd29, was found to have a defect in the initiation of the outer chain and displayed a temperature growth sensitivity at 37 degrees C allowing the isolation of the corresponding gene by complementation. Cloning, sequencing and disruption of NGD29 showed that it is a non lethal gene and identical to OCH1, It complemented both the glycosylation and growth defect, Membranes isolated from an ngd29 disruptant or an ngd29mnn1 double mutant were no longer able, in contrast to membranes from mild type cells, to transfer mannose from GDP mannose to Man(8)GlcNAc(2), the in vivo acceptor for building up the outer chain. Heterologous expression of glucose oxidase from Aspergillus niger in an ngd29mnn1 double mutant produced a secreted uniform glycoprotein with exclusively Man(8)GlcNAc(2) structure that in mild type yeast is heavily hyperglycosylated. The data indicate that this mutant strain is a suitable host for the expression of recombinant glycoproteins from different origin in S. cerevisiae to obtain mammalian oligomannosidic type N-linked carbohydrate chains.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | ASPARAGINE-LINKED OLIGOSACCHARIDES; PROTEIN GLYCOSYLATION; YEAST; ELONGATION; INVERTASE; EXPRESSION; DEFICIENT; CLONING; GLUCOSE; GENE; PROTEIN GLYCOSYLATION; GLYCOPROTEIN; GOLGI GLYCOSYLTRANSFERASE; SACCHAROMYCES CEREVISIAE |
| Depositing User: | Dr. Gernot Deinzer |
| Last Modified: | 19 Oct 2022 08:37 |
| URI: | https://pred.uni-regensburg.de/id/eprint/52386 |
Actions (login required)
 |
View Item |
