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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
46
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pubmed:dateCreated |
2001-11-12
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pubmed:abstractText |
Polymyxin-resistant mutants of Escherichia coli and Salmonella typhimurium accumulate a novel minor lipid that can donate 4-amino-4-deoxy-l-arabinose units (l-Ara4N) to lipid A. We now report the purification of this lipid from a pss(-) pmrA(C) mutant of E. coli and assign its structure as undecaprenyl phosphate-alpha-l-Ara4N. Approximately 0.2 mg of homogeneous material was isolated from an 8-liter culture by solvent extraction, followed by chromatography on DEAE-cellulose, C18 reverse phase resin, and silicic acid. Matrix-assisted laser desorption ionization/time of flight mass spectrometry in the negative mode yielded a single species [M - H](-) at m/z 977.5, consistent with undecaprenyl phosphate-alpha-l-Ara4N (M(r) = 978.41). (31)P NMR spectroscopy showed a single phosphorus atom at -0.44 ppm characteristic of a phosphodiester linkage. Selective inverse decoupling difference spectroscopy demonstrated that the undecaprenyl phosphate group is attached to the anomeric carbon of the l-Ara4N unit. One- and two-dimensional (1)H NMR studies confirmed the presence of a polyisoprene chain and a sugar moiety with chemical shifts and coupling constants expected for an equatorially substituted arabinopyranoside. Heteronuclear multiple-quantum coherence spectroscopy analysis demonstrated that a nitrogen atom is attached to C-4 of the sugar residue. The purified donor supports in vitro conversion of lipid IV(A) to lipid II(A), which is substituted with a single l-Ara4N moiety. The identification of undecaprenyl phosphate-alpha-l-Ara4N implies that l-Ara4N transfer to lipid A occurs in the periplasm of polymyxin-resistant strains, and establishes a new enzymatic pathway by which Gram-negative bacteria acquire antibiotic resistance.
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pubmed:grant |
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/4-amino-4-deoxyarabinose,
http://linkedlifedata.com/resource/pubmed/chemical/Amino Sugars,
http://linkedlifedata.com/resource/pubmed/chemical/Anti-Bacterial Agents,
http://linkedlifedata.com/resource/pubmed/chemical/Bacterial Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Carbohydrates,
http://linkedlifedata.com/resource/pubmed/chemical/DEAE-Cellulose,
http://linkedlifedata.com/resource/pubmed/chemical/Ethanolamines,
http://linkedlifedata.com/resource/pubmed/chemical/Lipid A,
http://linkedlifedata.com/resource/pubmed/chemical/Lipids,
http://linkedlifedata.com/resource/pubmed/chemical/Myristic Acids,
http://linkedlifedata.com/resource/pubmed/chemical/Palmitic Acid,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphorus,
http://linkedlifedata.com/resource/pubmed/chemical/Polymyxins,
http://linkedlifedata.com/resource/pubmed/chemical/Silicic Acid,
http://linkedlifedata.com/resource/pubmed/chemical/alpha-hydroxymyristic acid,
http://linkedlifedata.com/resource/pubmed/chemical/phosphorylethanolamine,
http://linkedlifedata.com/resource/pubmed/chemical/pmrA protein, Bacteria
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pubmed:status |
MEDLINE
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pubmed:month |
Nov
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pubmed:issn |
0021-9258
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
16
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pubmed:volume |
276
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
43132-44
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pubmed:dateRevised |
2007-11-15
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pubmed:meshHeading |
pubmed-meshheading:11535605-Amino Sugars,
pubmed-meshheading:11535605-Anti-Bacterial Agents,
pubmed-meshheading:11535605-Bacterial Proteins,
pubmed-meshheading:11535605-Carbohydrate Sequence,
pubmed-meshheading:11535605-Carbohydrates,
pubmed-meshheading:11535605-Cell Nucleus,
pubmed-meshheading:11535605-Cell-Free System,
pubmed-meshheading:11535605-Chromatography,
pubmed-meshheading:11535605-DEAE-Cellulose,
pubmed-meshheading:11535605-Escherichia coli,
pubmed-meshheading:11535605-Ethanolamines,
pubmed-meshheading:11535605-Hydrolysis,
pubmed-meshheading:11535605-Lipid A,
pubmed-meshheading:11535605-Lipids,
pubmed-meshheading:11535605-Magnetic Resonance Spectroscopy,
pubmed-meshheading:11535605-Models, Chemical,
pubmed-meshheading:11535605-Molecular Sequence Data,
pubmed-meshheading:11535605-Mutation,
pubmed-meshheading:11535605-Myristic Acids,
pubmed-meshheading:11535605-Palmitic Acid,
pubmed-meshheading:11535605-Periplasm,
pubmed-meshheading:11535605-Phosphorus,
pubmed-meshheading:11535605-Polymyxins,
pubmed-meshheading:11535605-Protein Binding,
pubmed-meshheading:11535605-Protein Conformation,
pubmed-meshheading:11535605-Protein Prenylation,
pubmed-meshheading:11535605-Salmonella typhimurium,
pubmed-meshheading:11535605-Silicic Acid,
pubmed-meshheading:11535605-Spectrometry, Mass, Matrix-Assisted Laser...
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pubmed:year |
2001
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pubmed:articleTitle |
Accumulation of a polyisoprene-linked amino sugar in polymyxin-resistant Salmonella typhimurium and Escherichia coli: structural characterization and transfer to lipid A in the periplasm.
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pubmed:affiliation |
Department of Biochemistry and the Duke NMR Spectroscopy Center, Duke University Medical Center, Durham, North Carolina 27710, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, U.S. Gov't, Non-P.H.S.,
Research Support, Non-U.S. Gov't
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