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Molecular Cloning and Expression of a Novel Cholinephosphotransferase Involved in Glycoglycerophospholipid Biosynthesis of Mycoplasma fermentans

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Abstract

A gene, mf1, encoding a novel cholinephosphotransferase in glycoglycerophospholipid (GGPL) biosynthesis of Mycoplasma fermentans PG18 was identified by genomic analysis, cloned, and expressed in Escherichia coli. The mf1 gene comprises an open reading frame of 777 bp encoding 258 amino acids. The mf1 gene product, Mf1, has 23% amino acid homology with LicD of Haemophilus influenzae but no homology with genes of other Mycoplasma species in the GenBank database. The reaction product of Mf1 using α-glucopyranosyl-1,2-dipalmitoilglycerol and cytidine 5′-diphosphocholine (CDP-choline) as substrates showed the specific protonated molecule at m/z 896, which corresponded to GGPL-I as determined by matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry (MALDI-TOF MS). Furthermore, the product ions of choline, phosphocholine, and hexose-bound phosphocholine were detected by tandem mass spectrometry (MS) analysis of protonated molecules at m/z 896. These results identified mf1 as a novel cholinephosphotransferase and showed that the phosphocholine transfer step is involved in the GGPL biosynthesis pathway of M. fermentans. This is the first report of a GGPL biosynthesis enzyme.

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References

  1. Dybvig K (1990) Mycoplasmal genetics. Annu Rev Microbiol 44:81–104

    Article  PubMed  CAS  Google Scholar 

  2. Haier J, Nasralla M, Franco AR et al (1999) Detection of mycoplasmal infections in blood of patients with rheumatoid arthritis. Rheumatology 38:504–509

    Article  PubMed  CAS  Google Scholar 

  3. Horowitz S, Evinson B, Borer A et al (2000) Mycoplasma fermentans in rheumatoid arthritis and other inflammatory arthritides. J Rheumatol 27:2747–2753

    PubMed  CAS  Google Scholar 

  4. Kawahito Y, Ichinose S, Sano H et al (2008) Mycoplasma fermentans glycolipid-antigen as a pathogen of rheumatoid arthritis. Biochem Biophys Res Commun 369:561–566

    Article  PubMed  CAS  Google Scholar 

  5. Kent C, Gee P, Lee SY et al (2004) A CDP-choline pathway for phosphatidylcholine biosynthesis in Treponema denticola. Mol Microbiol 51:471–481

    Article  PubMed  CAS  Google Scholar 

  6. Lysenko E, Richards JC, Cox AD et al (2000) The position of phosphorylcholine on the lipopolysaccharide of Haemophilus influenzae affects binding and sensitivity to C-reactive protein-mediated killing. Mol Microbiol 35:234–245

    Article  PubMed  CAS  Google Scholar 

  7. Maetinez-Morales F, Schobert M, Lopez-Lara IM et al (2003) Pathways for phosphatidylcholine biosynthesis in bacteria. Microbiology 149:3461–3471

    Article  Google Scholar 

  8. Matsuda K, Kasama T, Ishizuka I et al (1994) Structure of a novel phosphocholine-containing glycoglycerolipid from Mycoplasma fermentans. J Biol Chem 269:33123–33128

    PubMed  CAS  Google Scholar 

  9. Matsuda K, Ishizuka I, Kasama T et al (1997) Structure of a novel phosphocholine-containing aminoglycoglycerolipid of Mycoplasma fermentans. Biochim Biophys Acta 1349:1–12

    PubMed  CAS  Google Scholar 

  10. Matsuda K, Li JL, Harasawa R et al (1997) Phosphocholine-containing glycoglycerolipids (GGPL-I and GGPL-III) are species-specific major immunodeterminants of Mycoplasma fermentans. Biochem Biophys Res Commun 233:644–649

    Article  PubMed  CAS  Google Scholar 

  11. Nishida Y, Ohrui H, Meguro H et al (1994) Synthesis and absolute configuration of 6-o-phosphocholine-D-glucopyranosyl glycerolipid isolated from HTLV-I-infected cell lines. Tetrahedron Lett 35:5465–5468

    Article  CAS  Google Scholar 

  12. Sambook J, Russell W (2001) In vitro mutagenesis using double-stranded DNA templates: selection of mutants with DpnI. In: Molecular Cloning. vol. 2. Cold Spring Harbor, New York, pp. 13.19–13.25

  13. Schaeverbeke T, Gilroy CB, Bebear C et al (1996) Mycoplasma fermentans, but not M. penetrans, detected by PCR assays in synovium from patients with rheumatoid arthritis and other rheumatic disorders. J Clin Pathol 49:824–828

    Article  PubMed  CAS  Google Scholar 

  14. Sohlenkamp C, Lopez-Lara IM, Geoger O (2003) Biosynthesis of phosphatidylcholine in bacteria. Prog Lipid Res 42:115–162

    Article  PubMed  CAS  Google Scholar 

  15. Tamura K, Dudley J, Nei M et al (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evolution 24:1596–1599

    Article  CAS  Google Scholar 

  16. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties, and weight matrix choice. Nucleic Acids Res 22:4673–4680

    Article  PubMed  CAS  Google Scholar 

  17. Zhang JR, Heikkila II, Fischer W et al (1999) Pneumococcal licD2 gene is involved in phosphorylcholine metabolism. Mol Microbiol 31:1477–1488

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This research was supported by a grant-in-aid from the Ministry of Economy, Trade, and Industry of Japan.

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Authors and Affiliations

  1. Laboratory of Applied Microbiology, Graduate School of Agriculture, Hokkaido University, Kita-9, Nishi-9, Sapporo, 060-8589, Japan

    Noriko Ishida, Teruo Sone, Michiko Tanaka & Kozo Asano

  2. Department of Research and Developments, M Bio Technology Inc, Tokyo, Japan

    Daisuke Irikura, Kazuhiro Matsuda & Seiji Sato

Authors
  1. Noriko Ishida
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  2. Daisuke Irikura
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  3. Kazuhiro Matsuda
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  4. Seiji Sato
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  5. Teruo Sone
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  6. Michiko Tanaka
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  7. Kozo Asano
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Correspondence to Kozo Asano.

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Ishida, N., Irikura, D., Matsuda, K. et al. Molecular Cloning and Expression of a Novel Cholinephosphotransferase Involved in Glycoglycerophospholipid Biosynthesis of Mycoplasma fermentans . Curr Microbiol 58, 535–540 (2009). https://doi.org/10.1007/s00284-009-9362-6

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  • DOI: https://doi.org/10.1007/s00284-009-9362-6

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