Disruption of Cg-Ppml, a polyprenol monophosphomannose synthase, and the generation of lipoglycans-less mutants in Corynebacterium glutamicum

Kevin Gibson, L Eggeling, William Maughan, K Krumbach, Sudagar Gurcha, J Nigou, Gurdyal Besra

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

The glycosyl donor, polyprenyl monophosphomannose (PPM), has been shown to be involved in the biosynthesis of the mycobacterial lipoglycans: lipomannan and lipoarabinomannan. The mycobacterial PPM synthase (Mt-ppm1) catalyzes the transfer of mannose from GDP-mannose to polyprenyl phosphates. Based on sequence homology to Mt-ppm1, we have identified the PPM synthase from Corynebacterium glutamicum. In the present study, we demonstrate that the corynebacterial synthase is composed of two distinct domains; a catalytic domain (Cg-ppm1) and a membrane domain (Cg-ppm2). Through the inactivation of Cg-ppm1, we observed a complex phenotype that included altered cell growth rate and inability to synthesize PPM molecules and lipoglycans. When Cg-ppm2 was deleted, no observable phenotype was noted, indicating the clear organization of the two domains. The complementation of the inactivated Cg-ppm1 strain with the corresponding mycobacterial enzyme (Mt-Ppm1/D2) led to the restoration of a wild type phenotype. The present study illustrates, for the first time, the generation of a lipoglycan-less mutant based on a molecular strategy in a member of the Corynebacterianeae family. Lipoglycans are important immunomodulatory molecules involved in determining the outcome of infection, and so the generation of defined mutants and their subsequent immunological characterization is timely.
Original languageEnglish
Pages (from-to)40842-40850
Number of pages9
JournalJournal of Biological Chemistry
Volume278
Issue number42
Early online date31 Jul 2003
DOIs
Publication statusPublished - 31 Jul 2003

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