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Volume 163, Issue 2

Identification of lysophospholipase protein from and verification of its function Free

Abstract

is known to cause tremor disease in the Chinese mitten crab ; however, the molecular characterization of this pathogen is still unclear. has the ability to invade and survive within mouse 3T6 cells. The invasion process may require causing damage to the host cell membrane by chemical, physical or enzymatic means. In this study, we systematically characterized a novel lysophospholipase (lysoPL) of TDA-040725-5. The gene that encodes lysoPL in (SE-LysoPL) was cloned, sequenced and expressed in BL21 (DE3). Enzymatic assays revealed that the purified recombinant SE-LysoPL hydrolysed long-chain acyl esterases at pH 7 and 30 °C. SE-LysoPL was detected in the membrane and cytoplasmic protein fractions using the SE-LysoPL antibody in Western blot. The virulence ability of was effectively reduced at the early stage of infection (m.o.i.=100) by the SE-LysoPL antibody neutralization test. To the best of our knowledge, this is the first study to identify and characterize a gene from encoding a protein exhibiting lysoPL and esterase activities. Our findings indicate that SE-LysoPL plays important roles in the pathogenicity of .

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): enzyme activity , lysophospholipase , Spiroplasma eriocheiris and virulence factor
© 2017 The Authors
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2017-02-01
2024-04-18
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References

  1. Wang W, Chen J, Du K, Xu Z. Morphology of spiroplasmas in the Chinese mitten crab Eriocheir sinensis associated with tremor disease. Res Microbiol 2004; 155:630–635 [View Article][PubMed]
     [Google Scholar]
  2. Wang W, Gu W, Ding Z, Ren Y, Chen J et al. A novel Spiroplasma pathogen causing systemic infection in the crayfish Procambarus clarkii (Crustacea: Decapod), in China. FEMS Microbiol Lett 2005; 249:131–137 [View Article][PubMed]
     [Google Scholar]
  3. Wang W, Rong L, Gu W, Du K, Chen J. Study on experimental infections of Spiroplasma from the Chinese mitten crab in crayfish, mice and embryonated chickens. Res Microbiol 2003; 154:677–680 [View Article][PubMed]
     [Google Scholar]
  4. Wang W, Gu W, Gasparich GE, Bi K, Ou J et al. Spiroplasma eriocheiris sp. nov., associated with mortality in the Chinese mitten crab, Eriocheir sinensis. Int J Syst Evol Microbiol 2011; 61:703–708 [View Article][PubMed]
     [Google Scholar]
  5. Bi K, Huang H, Gu W, Wang J, Wang W. Phylogenetic analysis of Spiroplasmas from three freshwater crustaceans (Eriocheir sinensis, Procambarus clarkia and Penaeus vannamei) in China. J Invertebr Pathol 2008; 99:57–65 [View Article][PubMed]
     [Google Scholar]
  6. Regassa LB, Gasparich GE. Spiroplasmas: evolutionary relationships and biodiversity. Front Biosci 2006; 11:2983–3002 [View Article][PubMed]
     [Google Scholar]
  7. Razin S, Yogev D, Naot Y. Molecular biology and pathogenicity of mycoplasmas. Microbiol Mol Biol Rev 1998; 62:1094–1156[PubMed]
     [Google Scholar]
  8. Chambaud I, Wróblewski H, Blanchard A. Interactions between mycoplasma lipoproteins and the host immune system. Trends Microbiol 1999; 7:493–499 [View Article][PubMed]
     [Google Scholar]
  9. Gu W, Yao W, Zhao Y, Pei S, Jiang C et al. Establishment of spiroplasma-infected hemocytes as an in vitro laboratory culture model of Chinese mitten crab Eriocheir sinensis. Vet Microbiol 2014; 171:215–220 [View Article][PubMed]
     [Google Scholar]
  10. Yao W. Establishing Spiroplasma eriocheiris infect the two target cell model and screening interaction related proteins. Dissertation, Nanjing Normal University 2013
  11. Wang W, Zhu N, Li Z, Gu Z, Xu Z. Optical and transmission electron microscope study on nerve tissue infested by rickettsia-like organisms in crab, Eriocheir sinensis, suffering from tremor disease. Zool Res 2001; 22:467–471
     [Google Scholar]
  12. Wang W, Gu Z. Rickettsia-like organism associated with tremor disease and mortality of the Chinese mitten crab Eriocheir sinensis. Dis Aquat Organ 2002; 48:149–153 [View Article][PubMed]
     [Google Scholar]
  13. Ghannoum MA. Potential role of phospholipases in virulence and fungal pathogenesis. Clin Microbiol Rev 2000; 13:122–143 [View Article][PubMed]
     [Google Scholar]
  14. Nishizuka Y. Intracellular signaling by hydrolysis of phospholipids and activation of protein kinase C. Science 1992; 258:607–614 [View Article][PubMed]
     [Google Scholar]
  15. Weltzien HU. Cytolytic and membrane-perturbing properties of lysophosphatidylcholine. Biochim Biophys Acta 1979; 559:259–287 [View Article]
     [Google Scholar]
  16. Vernon LP, Bell JD. Membrane structure, toxins and phospholipase A2 activity. Pharmacol Ther 1992; 54:269–295 [View Article][PubMed]
     [Google Scholar]
  17. Golan DE, Brown CS, Cianci CM, Furlong ST, Caulfield JP. Schistosomula of Schistosoma mansoni use lysophosphatidylcholine to lyse adherent human red blood cells and immobilize red cell membrane components. J Cell Biol 1986; 103:819–828 [View Article][PubMed]
     [Google Scholar]
  18. Masamune A, Sakai Y, Satoh A, Fujita M, Yoshida M et al. Lysophosphatidylcholine induces apoptosis in AR42J cells. Pancreas 2001; 22:75–83[PubMed] [CrossRef]
     [Google Scholar]
  19. Zhou L, Shi M, Guo Z, Brisbon W, Hoover R et al. Different cytotoxic injuries induced by lysophosphatidylcholine and 7-ketocholesterol in mouse endothelial cells. Endothelium 2006; 13:213–226 [View Article][PubMed]
     [Google Scholar]
  20. Noverr MC, Cox GM, Perfect JR, Huffnagle GB. Role of PLB1 in pulmonary inflammation and cryptococcal eicosanoid production. Infect Immun 2003; 71:1538–1547 [View Article][PubMed]
     [Google Scholar]
  21. Wright LC, Payne J, Santangelo RT, Simpanya MF, Chen SC et al. Cryptococcal phospholipases: a novel lysophospholipase discovered in the pathogenic fungus Cryptococcus gattii. Biochem J 2004; 384:377–384 [View Article][PubMed]
     [Google Scholar]
  22. Wang W, Wen B, Gasparich GE, Zhu N, Rong L et al. A spiroplasma associated with tremor disease in the Chinese mitten crab (Eriocheir sinensis). Microbiology 2004; 150:3035–3040 [View Article][PubMed]
     [Google Scholar]
  23. Moulder RW, French FE, Chang CJ. Simplified media for spiroplasmas associated with tabanid flies. Can J Microbiol 2002; 48:1–6 [View Article][PubMed]
     [Google Scholar]
  24. Meng Q, Ou J, Ji H, Jiang X, Gu W et al. Identification and characterization of spiralin-like protein SLP25 from Spiroplasma eriocheiris. Vet Microbiol 2010; 144:473–477 [View Article][PubMed]
     [Google Scholar]
  25. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72:248–254 [View Article][PubMed]
     [Google Scholar]
  26. Wang J, Huang H, Feng Q, Liang T, Bi K et al. Enzyme-linked immunosorbent assay for the detection of pathogenic spiroplasma in commercially exploited crustaceans from China. Aquaculture 2009; 292:166–171 [CrossRef]
     [Google Scholar]
  27. Chu FH, Wang SY, Lee LC, Shaw JF. Identification and characterization of a lipase gene from Antrodia cinnamomea. Mycol Res 2008; 112:1421–1427 [View Article][PubMed]
     [Google Scholar]
  28. Salman M, Rottem S. The cell membrane of Mycoplasma penetrans: lipid composition and phospholipase A1 activity. Biochim Biophys Acta 1995; 1235:369–377 [View Article][PubMed]
     [Google Scholar]
  29. Kornspan JD, Rottem S. Phospholipase A and glycerophosphodiesterase activities in the cell membrane of Mycoplasma hyorhinis. FEMS Microbiol Lett 2012; 332:34–39 [View Article][PubMed]
     [Google Scholar]
  30. Duncan DB. Multiple range and multiple F tests. Biometrics 1955; 11:1–42 [CrossRef]
     [Google Scholar]
  31. Ku C, Lo WS, Chen LL, Kuo CH. Complete genomes of two dipteran-associated spiroplasmas provided insights into the origin, dynamics, and impacts of viral invasion in Spiroplasma. Genome Biol Evol 2013; 5:1151–1164 [View Article][PubMed]
     [Google Scholar]
  32. Nardini M, Dijkstra BW. Alpha/beta hydrolase fold enzymes: the family keeps growing. Curr Opin Struct Biol 1999; 9:732–737 [View Article][PubMed]
     [Google Scholar]
  33. Brenner S. The molecular evolution of genes and proteins: a tale of two serines. Nature 1988; 334:528–530 [View Article][PubMed]
     [Google Scholar]
  34. Liang T, Feng Q, Wu T, Gu W, Wang W. Use of oxytetracycline for the treatment of tremor disease in the Chinese mitten crab Eriocheir sinensis. Dis Aquat Organ 2009; 84:243–250 [View Article][PubMed]
     [Google Scholar]
  35. Schalk-Hihi C, Schubert C, Alexander R, Bayoumy S, Clemente JC et al. Crystal structure of a soluble form of human monoglyceride lipase in complex with an inhibitor at 1.35 Å resolution. Protein Sci 2011; 20:670–683 [View Article][PubMed]
     [Google Scholar]
  36. Sharp JD, Pickard RT, Chiou XG, Manetta JV, Kovacevic S et al. Serine 228 is essential for catalytic activities of 85-kDa cytosolic phospholipase A2. J Biol Chem 1994; 269:23250–23254[PubMed]
     [Google Scholar]
  37. Pickard RT, Chiou XG, Strifler BA, Defelippis MR, Hyslop PA et al. Identification of essential residues for the catalytic function of 85-kDa cytosolic phospholipase A2. Probing the role of histidine, aspartic acid, cysteine, and arginine. J Biol Chem 1996; 271:19225–19231 [View Article][PubMed]
     [Google Scholar]
  38. van Golde LM, Mcelhaney RN, van Deenen LL. A membrane-bound lysophospholipase from Mycoplasma laidlawii strain B. Biochim Biophys Acta 1971; 231:245–249 [View Article][PubMed]
     [Google Scholar]
  39. Gatt S, Morag B, Rottem S. Lysophospholipase-catalyzed hydrolysis of lysophospholipids in Mycoplasma gallisepticum membranes. J Bacteriol 1982; 151:1095–1101[PubMed]
     [Google Scholar]
  40. Snijder HJ, Ubarretxena-Belandia I, Blaauw M, Kalk KH, Verheij HM et al. Structural evidence for dimerization-regulated activation of an integral membrane phospholipase. Nature 1999; 401:717–721 [View Article][PubMed]
     [Google Scholar]
  41. Istivan TS, Coloe PJ. Phospholipase A in Gram-negative bacteria and its role in pathogenesis. Microbiology 2006; 152:1263–1274 [View Article][PubMed]
     [Google Scholar]
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Identification of lysophospholipase protein from Spiroplasma eriocheiris and verification of its function
Microbiology 163, 175 (2017); https://doi.org/10.1099/mic.0.000407
/content/journal/micro/10.1099/mic.0.000407
/content/journal/micro/10.1099/mic.0.000407
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