Sulfoquinovose synthase - an important enzyme in the N-glycosylation pathway of Sulfolobus acidocaldarius

Benjamin H. Meyer, Behnam Zolghadr, Elham Peyfoon, Martin Pabst, Maria Panico, Howard R. Morris, Stuart M. Haslam, Paul Messner, Christina Schäffer, More Authors

Research output: Contribution to journalArticleScientificpeer-review

56 Citations (Scopus)

Abstract

Recently, the Surface (S)-layer glycoprotein of the thermoacidophilic crenarchaeote Sulfolobus acidocaldarius was found to be N-glycosylated with a heterogeneous family of glycans, with the largest having a composition Glc 1Man 2GlcNAc 2 plus 6-sulfoquinovose. However, genetic analyses of genes involved in the N-glycosylation process in Crenarchaeota were missing so far. In this study we identify a gene cluster involved in the biosynthesis of sulfoquinovose and important for the assembly of the S-layer N-glycans. A successful markerless in-frame deletion of agl3 resulted in a decreased molecular mass of the S-layer glycoprotein SlaA and the flagellin FlaB, indicating a change in the N-glycan composition. Analyses with nanoLC ES-MS/MS confirmed the presence of only a reduced trisaccharide structure composed of Man 1GlcNAc 2, missing the sulfoquinovose, a mannose and glucose. Biochemical studies of the recombinant Agl3 confirmed the proposed function as a UDP-sulfoquinovose synthase. Furthermore, S.acidocaldarius cells lacking agl3 had a significantly lower growth rate at elevated salt concentrations compared with the background strain, underlining the importance of the N-glycosylation to maintain an intact and stable cell envelope, to enable the survival of S.acidocaldarius in its extreme environment.

Original languageEnglish
Pages (from-to)1150-1163
JournalMolecular Microbiology
Volume82
Issue number5
DOIs
Publication statusPublished - 2011
Externally publishedYes

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