Glycosylation of the phosphate binding protein, PstS, in Streptomyces coelicolor by a pathway that resembles protein O-mannosylation in eukaryotes

Silvia Wehmeier, Anpu Susan Varghese, S. S. Gurcha, B. Tissot, M. Panico, P. Hitchen, Ruth Morris, G. S. Besra, A. Dell, Margaret Caroline MacHin Smith

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Abstract

Previously mutations in a putative protein O-mannosyltransferase (SCO3154, Pmt) and a polyprenol phosphate mannose synthase (SCO1423, Ppm1) were found to cause resistance to phage, phi C31, in the antibiotic producing bacteria Streptomyces coelicolor A3(2). It was proposed that these two enzymes were part of a protein O-glycosylation pathway that was necessary for synthesis of the phage receptor. Here we provide the evidence that Pmt and Ppm1 are indeed both required for protein O-glycosylation. The phosphate binding protein PstS was found to be glycosylated with a trihexose in the S. coelicolor parent strain, J1929, but not in the pmt (-) derivative, DT1025. Ppm1 was necessary for the transfer of mannose to endogenous polyprenol phosphate in membrane preparations of S. coelicolor. A mutation in ppm1 that conferred an E218V substitution in Ppm1 abolished mannose transfer and glycosylation of PstS. Mass spectrometry analysis of extracted lipids showed the presence of a glycosylated polyprenol phosphate (PP) containing nine repeated isoprenyl units (C-45-PP). S. coelicolor membranes were also able to catalyse the transfer of mannose to peptides derived from PstS, indicating that these could be targets for Pmt in vivo.

Original languageEnglish
Pages (from-to)421-433
Number of pages13
JournalMolecular Microbiology
Volume71
Issue number2
Early online date11 Nov 2008
DOIs
Publication statusPublished - Jan 2009

Keywords

  • mycobacterium-tuberculosis
  • monophosphomannose synthase
  • corynebacterium-glutamicum
  • bacterial glycoproteins
  • Campylobacter jejuni
  • mannose synthase
  • system
  • A3(2)
  • smegmatis
  • lividans

Cite this

Wehmeier, S., Varghese, A. S., Gurcha, S. S., Tissot, B., Panico, M., Hitchen, P., Morris, R., Besra, G. S., Dell, A., & Smith, M. C. M. (2009). Glycosylation of the phosphate binding protein, PstS, in Streptomyces coelicolor by a pathway that resembles protein O-mannosylation in eukaryotes. Molecular Microbiology, 71(2), 421-433. https://doi.org/10.1111/j.1365-2958.2008.06536.x