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PMID: 20200052 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Protein tyrosine O-glycosylation--a rather unexplored prokaryotic glycosylation system.

Glycobiology ·Vol. 20 ·No. 6 ·2010-06-00 ·Pages 787-98

Zarschler K, Janesch B, Pabst M, Altmann F, Messner P, Schäffer C

Abstract

Glycosylation is a frequent and heterogeneous posttranslational protein modification occurring in all domains of life. While protein N-glycosylation at asparagine and O-glycosylation at serine, threonine or hydroxyproline residues have been studied in great detail, only few data are available on O-glycosidic attachment of glycans to the amino acid tyrosine. In this study, we describe the identification and characterization of a bacterial protein tyrosine O-glycosylation system. In the Gram-positive, mesophilic bacterium Paenibacillus alvei CCM 2051(T), a polysaccharide consisting of [-->3)-beta-d-Galp-(1[alpha-d-Glcp-(1-->6)] -->4)-beta-d-ManpNAc-(1-->] repeating units is O-glycosidically linked via an adaptor with the structure -[GroA-2-->OPO(2)-->4-beta-d-ManpNAc-(1-->4)] -->3)-alpha-l-Rhap-(1-->3)-alpha-l-Rhap-(1-->3)-alpha-l-Rhap-(1-->3)-beta-d-Galp-(1--> to specific tyrosine residues of the S-layer protein SpaA. A +AH4-24.3-kb S-layer glycosylation (slg) gene cluster encodes the information necessary for the biosynthesis of this glycan chain within 18 open reading frames (ORF). The corresponding translation products are involved in the biosynthesis of nucleotide-activated monosaccharides, assembly and export as well as in the transfer of the completed polysaccharide chain to the S-layer target protein. All ORFs of the cluster, except those encoding the nucleotide sugar biosynthesis enzymes and the ATP binding cassette (ABC) transporter integral transmembrane proteins, were disrupted by the insertion of the mobile group II intron Ll.LtrB, and S-layer glycoproteins produced in mutant backgrounds were analyzed by mass spectrometry. There is evidence that the glycan chain is synthesized in a process comparable to the ABC-transporter-dependent pathway of the lipopolysaccharide O-polysaccharide biosynthesis. Furthermore, with the protein WsfB, we have identified an O-oligosaccharyl:protein transferase required for the formation of the covalent beta-d-Gal-->Tyr linkage between the glycan chain and the S-layer protein.

MeSH Terms
Bacillus/chemistry,metabolism Bacterial Proteins/chemistry,genetics,metabolism Glycosylation Mutation Polysaccharides/chemistry,genetics,metabolism Reverse Transcriptase Polymerase Chain Reaction Tyrosine/analogs & derivatives,chemistry,metabolism
Chemicals
Bacterial Proteins Polysaccharides Tyrosine
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Zarschler Kristof
Department of NanoBiotechnology, ViennaInstitute of BioTechnology, Universität für Bodenkultur Wien, A-1190 Vienna,Austria.
Janesch Bettina
Pabst Martin
Altmann Friedrich
Messner Paul
Schäffer Christina
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Article Info
Journal
Glycobiology
Abbr.
Glycobiology
ISSN
1460-2423
DOI
10.1093/glycob/cwq035
Published
2010-06-00
Epub
2010-00-03
Pages
787-98
Language
English
Region
England
NLM ID
9104124
PMCID
PMC4397588
Subset
IM
Grants
Austrian Science Fund FWF · P 19047 · Austria
Austrian Science Fund FWF · P 20745 · Austria
External Links
PubMed source DOI: 10.1093/glycob/cwq035 PMC Full Text View journal details
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