Comprehensive structure characterization of lipid a extracted from Yersinia pestis for determination of its phosphorylation configuration View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2010-01-25

AUTHORS

Jace W. Jones, Ilana E. Cohen, František Tureĉek, David R. Goodlett, Robert K. Ernst

ABSTRACT

We report on comprehensive structure characterization of lipid A extracted from Yersinia pestis (Yp) for determination of its phosphorylation configuration that was achieved by combining the methods of molecular biology with high-resolution tandem mass spectrometry. The phosphorylation pattern of diphosphorylated lipid A extracted from Yp has recently been found to be a heterogeneous mixture of C-1 and C-4′ bisphosphate, C-1 pyrophosphate, and C-4′ pyrophosphate (Proc. Natl. Acad. Sci.2008,105, 12742). To reduce the inherent phosphate heterogeneity of diphosphorylated lipid A extracted from Yp, we incorporated specific C-1 and C-4′ position phosphatases into wild type KIM6+ Yp grown at 37°C. Comprehensive high-resolution tandem mass spectrometric analyses of lipid A extracted from Yp modified with either the C-1 or C-4′ phosphatase allowed for unambiguous structure assignment of monophosphorylated and diphosphorylated lipid A and distinction of isomeric bisphosphate and pyrophosphate forms. The prevalent aminoarabinose modification was determined to be exclusively attached to the lipid A disaccharide via a phospho-diester linkage at either or both the C-1 and C-4′ positions. More... »

PAGES

785-799

Identifiers

URI

http://scigraph.springernature.com/pub.10.1016/j.jasms.2010.01.008

DOI

http://dx.doi.org/10.1016/j.jasms.2010.01.008

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1003632008

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/20185334


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