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Poster Presentations
Day 1, June 10（Mon.） Room P1 (Multipurpose Hall)・Room P2 (Conference Room 101+102)
- 1P-37
Quantification of Polyprenyl Diphosphates in Escherichia coli Cells Using High-performance Liquid Chromatography
(1Toho Univ., 2Wakayama Med. Univ.)
Tomotaka Jitsukawa1, Soichiro Watanabe1, oYasushi Shigeri2, Shingo Fujisaki1
Although dephosphorylation of undecaprenyl diphosphate is considered important in regulating the biosynthesis of undecaprenyl phosphate, the levels of these substances have not been measured separately. We have developed a method for the measurement of the intracellular levels of these substances. Polyprenyl phosphates and diphosphates prepared by chemical phosphorylation of polyprenols from Staphylococcus aureus were used to establish the conditions for fractionation by ion-exchange chromatography and high-performance liquid chromatography (HPLC). By using an elution solvent containing tetraethylammonium phosphate as an ion-pair reagent for HPLC, polyprenyl phosphate and polyprenyl diphosphate with carbon numbers from 40 to 55 could be detected as separate peaks from the reversed-phase column. This analytical method was applied to lipids extracted from Escherichia coli to determine the intracellular levels of octaprenyl phosphate, undecaprenyl phosphate, octaprenyl diphosphate, and undecaprenyl diphosphate. This is the first report of separate measurement of cellular levels of polyprenyl phosphates and polyprenyl diphosphates.