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J Biol Chem, Vol. 275, Issue 2, 937-941, January 14, 2000

Structural Characterization of the Inflammatory Moiety of a Variable Major Lipoprotein of Borrelia recurrentis*

Ian G. ScraggDagger , Dominic KwiatkowskiDagger §, and Vincent VidalDagger

From the Dagger  University Department of Paediatrics, John Radcliffe Hospital, Oxford OX3 9DU, United Kingdom

Andrew Reason, Thanai Paxtonpar , Maria Panicopar , Ann Dellpar , and Howard Morrispar

From  M-Scan, Silwood Park, Ascot SL5 7PZ, United Kingdom and the par  Department of Biochemistry, Imperial College, London SW7 2AZ, United Kingdom

Louse-borne relapsing fever, caused by Borrelia recurrentis, provides one of the best documented examples of the causative role of tumor necrosis factor (TNF) in the pathology of severe infection in humans. We have identified the principal TNF-inducing factor of B. recurrentis as a variable major lipoprotein (Vmp). Here we report the complete gene sequence of Vmp, including its lipoprotein leader sequence. Using metabolically labeled forms of the native Vmp we confirm that the TNF inducing properties are associated with the lipid portion of the molecule. Quadrupole orthogonal time of flight mass spectrometry unequivocally locates the lipidic moiety at the NH2-terminal cysteine of the native polypeptide, and indicates the existence of three forms which are consistent with the structures C16:0, C16:0, C16:0 glyceryl cysteine; C18:1, C16:0, C16:0 glyceryl cysteine; and C18:0, C16:0, C16:0 glyceryl cysteine. These data provide the first direct evidence that the TNF inducing lipid modification of native Borrelia lipoproteins is a structural homologue of the murein lipoprotein of Escherichia coli.

* This work was supported by the Medical Research Council (United Kingdom) and the quadrupole orthogonal time of flight research was funded by the Wellcome Trust (to H. R. M. and A. D.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ To whom correspondence should be addressed: Dept. of Paediatrics, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DU, United Kingdom. Tel.: 44-1865-221071; Fax: 44-1865-220479; E-mail: dominic.kwiatkowski@paediatrics.ox.ac.uk.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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