Extraction and identification by mass spectrometry of undecaprenyl diphosphate-MurNAc-pentapeptide-GlcNAc from Escherichia coli.
Undecaprenyl diphosphate-MurNAc-pentapeptide-GlcNAc (lipid II) is extracted from Escherichia coli cells by utilizing its unusual pH-dependent solubility property in a Bligh-Dyer system, and identified by electrospray ionization mass spectrometry in conjunction with a novel 15N mass shift analysis. The described approach will facilitate the structural characterization of lipid II variants from diverse bacteria, including antibiotic-resistant mutants, as well as the numerous minor uncharacterized lipids present in all biological systems.
Guan
Ziqiang
Z
Department of Biochemistry, Duke University Medical Center, Durham, NC 27710, USA. zguan@biochem.duke.edu
Breazeale
Steven D
SD
Raetz
Christian R H
CR
D011756:Diphosphates / Q000737:chemistry*; D004926:Escherichia coli / Q000254:growth & development / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D013058:Mass Spectrometry; D009112:Muramic Acids / Q000737:chemistry*; D012995:Solubility; D021241:Spectrometry, Mass, Electrospray Ionization
D011756:Diphosphates; D009112:Muramic Acids