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Microbiology 153 (2007), 2621-2629; DOI  10.1099/mic.0.2007/008078-0
© 2007 Society for General Microbiology

Structural characterization of a partially arabinosylated lipoarabinomannan variant isolated from a Corynebacterium glutamicum ubiA mutant

Raju Venkata Veera Tatituri1, Luke J. Alderwick1, Arun K. Mishra1, Jerome Nigou2, Martine Gilleron2, Karin Krumbach3, Paul Hitchen4, Assunta Giordano5, Howard R. Morris4,6, Anne Dell4, Lothar Eggeling3 and Gurdyal S. Besra1

1 School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK
2 Institut de Pharmacologie et de Biologie Structurale, UMR CNRS 5089, Toulouse, France
3 Institute for Biotechnology 1, Research Centre Juelich, D-52425 Juelich, Germany
4 Division of Molecular Biosciences, Faculty of Natural Sciences, Imperial College, London SW7 2AZ, UK
5 Istituto di Chimica Biomolecolare, Consiglio Nazionale delle Ricerche, 80078 Pozzuoli (NA), Italy
6 M-Scan Ltd., Wokingham RG41 2TZ, UK

Correspondence
Gurdyal S. Besra
g.besra{at}bham.ac.uk

Arabinan polysaccharide side-chains are present in both Mycobacterium tuberculosis and Corynebacterium glutamicum in the heteropolysaccharide arabinogalactan (AG), and in M. tuberculosis in the lipoglycan lipoarabinomannan (LAM). This study shows by quantitative sugar and glycosyl linkage analysis that C. glutamicum possesses a much smaller LAM version, Cg-LAM, characterized by single t-Araf residues linked to the {alpha}(1->6)-linked mannan backbone. MALDI-TOF MS showed an average molecular mass of 13 800–15 400 Da for Cg-LAM. The biosynthetic origin of Araf residues found in the extracytoplasmic arabinan domain of AG and LAM is well known to be provided by decaprenyl-monophosphoryl-D-arabinose (DPA). However, the characterization of LAM in a C. glutamicum : : ubiA mutant devoid of prenyltransferase activity and devoid of DPA-dependent arabinan deposition into AG revealed partial formation of LAM, albeit with a slightly altered molecular mass. These data suggest that in addition to DPA utilization as an Araf donor, alternative pathways exist in Corynebacterianeae for Araf delivery, possibly via an unknown sugar nucleotide.

Abbreviations: Ac, acylated; Ara, arabinose; Araf, arabinofuranose; AG, arabinogalactan; DPA, decaprenyl-monophosphoryl-D-arabinose; DPM, decaprenyl-monophosphoryl-mannose; DPR, decaprenyl-monophosphoryl-D-ribose; DPPR, decaprenyl-monophosphoryl-5-phosphoribose; LAM, lipoarabinomannan; LM, lipomannan; Man, mannose; mAGP, mycolyl-arabinogalactan-peptidoglycan; p, pyranose; PI, phosphatidyl-myo-inositol; PIMs, phosphatidyl-myo-inositol mannosides; pRpp, 5-phosphoribofuranose pyrophosphate; t-, terminal; TFA, trifluoroacetic acid




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D. J. Lea-Smith, K. L. Martin, J. S. Pyke, D. Tull, M. J. McConville, R. L. Coppel, and P. K. Crellin
Analysis of a New Mannosyltransferase Required for the Synthesis of Phosphatidylinositol Mannosides and Lipoarbinomannan Reveals Two Lipomannan Pools in Corynebacterineae
J. Biol. Chem., March 14, 2008; 283(11): 6773 - 6782.
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