TY - JOUR
T1 - Sulfoquinovose synthase - an important enzyme in the N-glycosylation pathway of Sulfolobus acidocaldarius
AU - Meyer, Benjamin H.
AU - Zolghadr, Behnam
AU - Peyfoon, Elham
AU - Pabst, Martin
AU - Panico, Maria
AU - Morris, Howard R.
AU - Haslam, Stuart M.
AU - Messner, Paul
AU - Schäffer, Christina
AU - More Authors, null
PY - 2011
Y1 - 2011
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=82155175626&partnerID=8YFLogxK
U2 - 10.1111/j.1365-2958.2011.07875.x
DO - 10.1111/j.1365-2958.2011.07875.x
M3 - Article
C2 - 22059775
AN - SCOPUS:82155175626
SN - 0950-382X
VL - 82
SP - 1150
EP - 1163
JO - Molecular Microbiology
JF - Molecular Microbiology
IS - 5
ER -