Exchange of Ser-4 for Val, Leu or Asn in the sequon Asn-Ala-Ser does not prevent N-glycosylation of the cell surface glycoprotein from Halobacterium halobium

Zeitler, Reinhard and Hochmuth, Eduard and Deutzmann, Rainer and Sumper, Manfred (1998) Exchange of Ser-4 for Val, Leu or Asn in the sequon Asn-Ala-Ser does not prevent N-glycosylation of the cell surface glycoprotein from Halobacterium halobium. GLYCOBIOLOGY, 8 (12). pp. 1157-1164. ISSN 0959-6658, 1460-2423

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Abstract

The archaeon Halobacterium halobium expresses a cell surface glycoprotein (CSG) with a repeating pentasaccharide unit N-glycosidically linked via N-acetylgalactosamine to Asn-2 of the polypeptide (GalNAc(1-N)Asn linkage type). This asparagine of the linkage unit is located within the N-terminal sequence Ala-Asn-Ala-Ser-, in accordance with the tripeptide consensus sequence Asn-Xaa-Ser/Thr typical for nearly every N-glycosylation site known so far, which are of the GlcNAc(1-N)-Asn linkage type. By a gene replacement method csg mutants were created which replace the serine residue of the consensus sequence by valine, leucine, and asparagine. Unexpectedly, this elimination of the consensus sequence did not prevent N-glycosylation. All respective mutant cell surface glycoproteins were N-glycosylated at Asn-2 with the same N-glycan chain as the wild type CSG, Asn-479 is N-glycosylated via a Glc(1-N)Asn linkage type in the wild type CSG. Replacement of Ser-481 in the sequence Asn-Ser-Ser for valine prevented glycosylation of Asn-479. From these results we postulate the existence of two different N-glycosyltransferases in H.halobium, one of which does not use the typical consensus sequence Asn-Xaa-Ser/Thr necessary for all other N-glycosyltransferases described so far.

Item Type: Article
Uncontrolled Keywords: HYDROXY AMINO-ACID; WALL GLYCOPROTEIN; SHUTTLE VECTORS; BACTERIO-OPSIN; LINKAGE UNIT; GENE; EXPRESSION; ARCHAEBACTERIUM; BIOSYNTHESIS; SEQUENCE; archaea; gene replacement; N-glycosylation; halobacteria
Subjects: 500 Science > 540 Chemistry & allied sciences
500 Science > 570 Life sciences
Divisions: Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Alumni or Retired > Prof. Dr. Manfred Sumper
Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie I > Prof. Dr. Rainer Deutzmann
Depositing User: Dr. Gernot Deinzer
Date Deposited: 09 Feb 2023 07:46
Last Modified: 09 Feb 2023 07:46
URI: https://pred.uni-regensburg.de/id/eprint/49292

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