Abstract
Bacteriovorax stolpii is a predator of larger gram-negative bacteria and lives as a parasite in the intraperiplasmic space of the host cell. This bacterium is unusual among prokaryotes in that sphingolipids comprise a large proportion of its lipids. We here report the presence of 18 molecular species of B. stolpii UKi2 sphingophosphonolipids (SPNLs). 31P NMR spectroscopy and analysis of Pi released by a differential hydrolysis protocol confirmed the phosphonyl nature of these lipids. The SPNLs were dominated by those with 1-hydroxy-2-aminoethane phosphonate (hydroxy-aminoethylphosphonate) polar head groups; aminoethylphosphonate was also detected in minor SPNL components. The long-chain bases (LCBs) were dominated by C17 iso-branched phytosphingosine; C17 iso-branched dihydrosphingosine was also present in some SPNLs. The N-linked fatty acids were predominantly iso-branched and most contained an α-hydroxy group (C15 α-hydroxy fatty acid was the major fatty acid). Minor molecular species containing nonhydroxy fatty acids were also detected. The definitive iso-structures of the predominant fatty acids and LCBs present in the B. stolpii SPNLs were established using 13C and 3H nuclear magnetic resonance spectroscopy; less than 20% were unbranched. Detection and analyses of intact compounds by MS-MS were performed by a hybrid quadrupole time-of-flight (Q-TOF-II) MS equipped with an electrospray ionization source. Analyses of peracetylated derivatives verified the structural assignments of these lipids.
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Baer, M. L.; Ravel, J.; Chun, J.; Hill, R. T.; Williams, H. N. A Proposal for the Reclassification of Bdellovibrio stolpii and Bdellovibrio starrii into a New Genus, Bacteriovorax. gen. nov. as Bacteriovorax stolpii comb. nov. and Bacteriovorax starrii comb. nov., respectively. Int. J. Syst. Evol. Microbiol. 2000, 50, 219–224.
Hespell, R. B.; Miozzari, G. F.; Rittenberg, S. C. Ribonucleic Acid Destruction and Synthesis during Intraperiplasmic Growth of Bdellovibrio bacteriovorus. J. Bacteriol. 1975, 123, 481–491.
Kuenen, J. G.; Rittenberg, S. C. Incorporation of Long-Chain Fatty Acids of the Substrate Organism by Bdellovibrio bacteriovorax during Intraperiplasmic Growth. J. Bacteriol. 1975, 121, 1145–1157.
Steiner, S.; Conti, S. F.; Lester, R. L. Occurrence of Phosphonosphingolipids in Bdellovibrio bacteriovorus Strain Uki2. J. Bacteriol. 1973, 116, 1199–1211.
Stoffel, W. Sphingolipids. Ann. Rev. Biochem. 1971, 40, 57–82.
Morrel, P.; Braun, B. Biosynthesis and Metabolic Degradation of Sphingolipids Not Containing Sialic Acid. J. Lipid Res. 1972, 13, 293–309.
Ferguson, K. A.; Conner, R. L.; Mallory, F. B.; Mallory, C. W. α-Hydroxy Fatty Acids in Sphingolipids of Tetrahymena. Biochim. Biophys. Acta. 1972, 270, 111–116.
Viswanathan, C. V.; Rosenberg, H. Isolation of Ceramide Monomethylaminoethylphosphonate from the Lipids of Tetrahymena pyriformis W. J. Lipid Res. 1973, 14, 327–330.
Ferguson, K. A.; Davis, F. M.; Conner, R. L.; Landrey, J. R.; Mallory, F. B. Effect of Sterol Replacement in Vivo on the Fatty Acid Composition of Tetrahymena. J. Biol. Chem. 1975, 250, 6998–7005.
Sugita, M.; Fukunaga, Y.; Ohkawa, K.; Nozawa, Y.; Hori, T. Structural Components of Sphingophosphonolipids from the Ciliated Protozoan, Tetrahymena pyriformis WH-14. J. Biochem. (Tokyo). 1979, 86, 281–288.
Rhoads, D. E.; Kaneshiro, E. S. Characterizations of Phospholipids from Paramecium tetraurelia Cells and Cilia. J. Protozool. 1979, 26, 329–338.
Kaneshiro, E. S.; Matesic, D. F.; Jayasimhulu, K. Characterizations of Six Ethanolamine Sphingophospholipids from Paramecium Cells and Cilia. J. Lipid Res. 1984, 25, 369–377.
Kaneshiro, E. S.; Jayasimhulu, K.; Lester, R. L. Characterizations of Inositol Lipids from Leishmania donovani Promastigotes: Identification of an Inositol Sphingolipid. J. Lipid Res. 1986, 27, 1294–1303.
Kaneshiro, E. S.; Jayasimhulu, K.; Sul, D.; Erwin, J. A. Identification and Initial Characterizations of Free, Glycosylated, and Phosphorylated Ceramides of Paramecium. J. Lipid Res. 1997, 38, 2399–2410.
Kariotoglou, D. M.; Mastronicolis, S. K. Sphingophosphonolipid Molecular Species from Edible Mollusks and a Jellyfish. Comp. Biochem. Physiol. B Biochem. Mol. Biol. 2003, 136, 27–44.
Liu, G.; Kleine, L.; Hebert, R. L. Advances in the Signal Transduction of Ceramide and Related Sphingolipids. Crit. Rev. Clin. Lab. Sci. 1999, 36, 511–573.
Hannun, Y. A.; Luberto, C. Ceramide in the Eukaryotic Stress Response. Trends Cell. Biol. 2000, 10, 73–80.
Futerman, A. H.; Hannun, Y. A. The Complex Life of Simple Sphingolipids. EMBO Rep. 2004, 5, 777–782.
Pettus, B. J.; Chalfant, C. E.; Hannun, Y. A. Sphingolipids in Inflammation: Roles and Implications. Curr. Mol. Med. 2004, 4, 405–418.
Brown, D. A.; London, E. Structure and Function of Sphingolipid- and Cholesterol-Rich Membrane Rafts. J. Biol. Chem. 2000, 275, 17221–17224.
London, E.; Brown, D. A. Insolubility of Lipids in Triton X-100: Physical Origin and Relationship to Sphingolipid/Cholesterol Membrane Domains (Rafts). Biochim. Biophys. Acta. 2000, 1508L, 182–195.
Chazal, N.; Gerlier, D. Virus Entry, Assembly, Budding, and Membrane Rafts. Microbiol. Mol. Biol. Rev. 2003, 67, 226–237.
Gulbins, E.; Dreschers, S.; Wilder, B.; Grassme, H. Ceramide, Membrane Rafts, and Infections. J. Mol. Med. 2004, 82, 357–363.
Martin, S. W.; Konopka, J. B. Lipid Raft Polarization Contributes to Hyphal Growth in Candida albicans. Eukaryot. Cell. 2004, 3, 675–684.
Seveau, S.; Bierne, H.; Giroux, S.; Prevost, M. C.; Cossart, P. Role of Lipid Rafts in E-Cadherin- and HGF-R/Met-Mediated Entry of Listeria monocytogenes into Host Cells. J. Cell Biol. 2004, 166, 743–753.
van Meer, G.; Sprong, H. Membrane Lipids and Vesicular Traffic. Curr. Opin. Cell Biol. 2004, 16, 373–378.
Kawahara, K.; Seydel, U.; Matsuura, M.; Danbara, H.; Rietschel, E. T.; Zahringer, U. Chemical Structure of Glycosphingolipids Isolated from Sphingomonas paucimobilis. FEBS Lett. 1991, 292, 107–110.
LaBach, J. P.; White, D. C. Identification of Phosphorylethanolamine and Ceramide Phosphorylglycerol in the Lipids of an Anaerobic Bacterium. J. Lipid Res. 1969, 10, 528–534.
Watanabe, Y.; Nakajima, M.; Hoshino, T.; Jayasimhulu, K.; Brooks, E. E.; Kaneshiro, E. S. A Novel Sphingophosphonolipid Head Group 1-Hydroxy-2-Aminoethyl Phosphonate in Bdellovibrio stolpii. Lipids 2001, 36, 513–519.
Sugita, M.; Hori, T. Isolation of Diacylglycerol-2-Aminoethylphosphonate from Tetrahymena pyriformis. J. Biochem. (Tokyo) 1971, 69, 1149–1150.
Matesic, D. F.; Kaneshiro, E. S. Incorporation of Serine into Paramecium Ethanolamine Phospholipid and Phosphonolipid Head Groups. Biochem. J. 1984, 222, 229–233.
Kaneshiro, E. S. Lipids of Paramecium. J. Lipid Res. 1987, 28, 1241–1258.
Liebisch, G.; Drobnik, W.; Reil, M.; Trumbach, B.; Arnecke, R.; Olgemoller, B.; Roscher, A.; Schmitz, G. Quantitative Measurement of Different Ceramide Species from Crude Cellular Extracts by Electrospray Ionization Tandem Mass Spectrometry (ESI-MS/MS). J. Lipid Res. 1999, 40, 1539–1546.
Costello, C. E.; Beach, D. H.; Singh, B. N. Acidic Glycerol Lipids of Trichomonas vaginalis and Tritrichomonas foetus. Biol. Chem. 2001, 382, 275–282.
Jensen, N. J.; Tomer, K. B.; Gross, M. L. FAB MS/MS for Phosphatidylinositol, -glycerol, -ethanolamine and Other Complex Phospholipids. Lipids 1987, 22, 480–409.
Bligh, E. G.; Dyer, W. J. A Rapid Method of Total Lipid Extraction and Purification. Can. Biochem. Physiol. 1959, 37, 911–917.
Dittmer, J. C.; Lester, R. L. A Simple, Specific Spray for the Detection of Phospholipids on Thin-Layer Chromatograms. J Lipid Res. 1964, 15, 126–127.
Harrison, K. A.; Davies, S. S.; Marathe, G. K.; McIntyre, T.; Prescott, S.; Reddy, K. M.; Falck, J. R.; Murphy, R. C. Analysis of Oxidized Glycerophosphocholine Lipids Using Electrospray Ionization Mass Spectrometry and Microderivatization Techniques. J. Mass Spectrom. 2000, 35, 224–236.
Glonek, T.; Henderson, T. O.; Hilderbrand, R. L.; Myers, T. C. Biological Phosphonates: Determination by Phosphorus-31 Nuclear Magnetic Resonance. Nature 1970, 169, 192–194.
Meneses, P.; Glonek, T. High Resolution 31P NMR of Extracted Phospholipids. J. Lipid Res. 1988, 29, 679–687.
Liang, C. R.; Rosenberg, H. The Biosynthesis of the Carbon-Phosphorus Bond in Tetrahymena. Biochim. Biophys. Acta. 1968, 156, 437–439.
Rosenberg, H. Phosphonolipids. In Form and Function of Phospholipids; Ansell, G. B.; Dawson, R. M. C.; Hawthorne, J. N., Eds.; Elsevier Publishing: Amsterdam, 1973; pp 333–344.
Hori, T.; Nozawa, Y. Phosphonolipids. In Phospholipids; Hawthorne, J. N.; Ansell, E. G., Eds.; Elsevier Biomedical Press: New York, 1982; pp 95–128.
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Published online November 20, 2006
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Jayasimhulu, K., Hunt, S.M., Kaneshiro, E.S. et al. Detection and identification of Bacteriovorax stolpii UKi2 sphingophosphonolipid molecular species. J Am Soc Mass Spectrom 18, 394–403 (2007). https://doi.org/10.1016/j.jasms.2006.10.014
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DOI: https://doi.org/10.1016/j.jasms.2006.10.014