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. 1985 Mar;161(3):861–867. doi: 10.1128/jb.161.3.861-867.1985

Role of lipid intermediate(s) in the synthesis of serogroup B Neisseria meningitidis capsular polysaccharide.

L Masson, B E Holbein
PMCID: PMC214976  PMID: 3918990

Abstract

Neisseria meningitidis serogroup B strain M986 was examined for the involvement of lipid intermediate(s) participating in the biosynthesis of the sialic acid capsular polysaccharide. The addition of exogenous undecaprenyl phosphate, phosphatidylethanolamine, or phosphatidylglycerol to particulate membranes, in the presence of cytidine 5'-monophosphosialic acid, resulted in the stimulation of sialyltransferase activity specifically by undecaprenyl phosphate. Sialyltransferase activity, after delipidation of particulate membrane proteins, was specifically reconstituted by undecaprenyl phosphate. After the addition of 14C-labeled cytidine 5'-monophosphosialic acid to particulate membranes, the level of labeled lipid intermediate(s), extracted by chloroform-methanol (2:1), increased up to a maximum level between 3.75 and 5.0 min, which subsequently decreased to a lower steady-state level. Pulse-chase experiments revealed a transient, solvent-extractable, lipid-linked component. The extracted N-acetylneuraminic acid was in polymeric form. Sequential oxidation and reduction of the extracted radioactivity followed by neuraminidase treatment revealed an average degree of polymerization of four or five N-acetylneuraminic acid residues. Bacitracin-sensitive peptidoglycan was synthesized in vitro by particulate membranes. Cross-competition experiments between peptidoglycan and capsular polysaccharide synthesis by preincubation of precursors of one pathway during synthesis of the other revealed a competitive effect for a common component. This component was believed to be a common pool of undecaprenyl phosphate. A model for the production and regulation of the capsular polysaccharide is proposed.

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Selected References

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