Skip to main content

Advertisement

SpringerLink
Log in

Molecular characterization and serodiagnosis analysis of a novel lysophospholipase from Clonorchis sinensis

  • Original Paper
  • Published:
Parasitology Research Aims and scope Submit manuscript

Abstract

A cDNA clone encoding a novel lysophospholipase with a predicted molecular weight of 25.2 kDa was isolated from a Clonorchis sinensis adult cDNA library. The enzyme activity of the recombinant protein expressed in Escherichia coli was determined using phosphatidylcholine and lysophosphatidylcholine as substrates. Western blotting analysis indicated that it belonged to excretory/secretory proteins of the adults. The sensitivity and specificity of the recombinant antigen for serodiagnosis were evaluated with immunoglobulin enzyme-linked immunosorbent assay using serum samples from 20 patients with clonorchiasis and 20 patients with schistosomiasis. The sensitivity (75%) and specificity (80%) of the recombinant protein were comparable to those of crude extracts, at 65 and 82.5%, respectively. The sensitivity of the recombinant protein was 77% using 100 serum samples of clonorchiasis patients with various parasite burden. The results suggested that the recombinant lysophospholipase protein was not a satisfactory candidate for diagnosis of clonorchiasis, although it might be an excretory/secretory protein.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Berrizbeitia M, Ndao M, Bubis J, Gottschalk M, Ache A, Lacouture S, Medina M, Ward BJ (2006) Purified excreted–secreted antigens from Trypanosoma cruzi trypomastigotes as tools for diagnosis of Chagas’ disease. J Clin Microbiol 44:291–296

    Article  PubMed  CAS  Google Scholar 

  • Bhanot P, Schauer K, Coppens I, Nussenzweig V (2005) A surface phospholipase is involved in the migration of plasmodium sporozoites through cells. J Biol Chem 280:6752–6760

    Article  PubMed  CAS  Google Scholar 

  • Brenner S (1988) The molecular evolution of genes and proteins: a tale of two serines. Nature 334:528–530

    Article  PubMed  CAS  Google Scholar 

  • Cervi L, Rossi G, Masih DT (1999) Potential role for excretory–secretory forms of glutathione-S-transferase (GST) in Fasciola hepatica. Parasitology 119:627–633

    Article  PubMed  CAS  Google Scholar 

  • Chen SCA, Wright LC, Golding JC, Sorrell TC (2000) Purification and characterization of secretory phospholipase B, lysophospholipase and lysophospholipase/transacylase from a virulent strain of the pathogenic fungus Cryptococcus neoformans. Biochem J 347:431–439

    Article  PubMed  CAS  Google Scholar 

  • Choi MH, Park IC, Li S, Hong ST (2003) Excretory–secretory antigen is better than crude antigen for the serodiagnosis of clonorchiasis by ELISA. Korean J Parasitol 41:35–39

    PubMed  Google Scholar 

  • Chung YY, Ko RC (1999) A novel cDNA clone encoding a specific excretory/secretory antigen of larval Trichinella pseudospiralis. Parasitol Res 85:685–691

    Article  PubMed  CAS  Google Scholar 

  • de Oliveira D, de Souza Tde A, Murate LS, Jankevicius JV, Gaziri LC, Jankevicius SI (2004) Protease and phospholipase inhibition protect Veneza zonata (Hemiptera Coreidae) against septicemia caused by parasite trypanosomatid 563DT. J Invertebr Pathol 85:9–17

    Article  PubMed  CAS  Google Scholar 

  • Djordjevic JT, Del Poeta M, Sorrell TC, Turner KM, Wright LC (2005) Secretion of cryptococcal phospholipase B1 (PLB1) is regulated by a glycosylphosphatidylinositol (GPI) anchor. Biochem J 389:803–812

    Article  PubMed  CAS  Google Scholar 

  • Ganga G, Varshney JP, Sharma RL (2006) Effect of Fasciola gigantica excretory secretory antigen on rat hematological indices. J Vet Sci 7:123–125

    PubMed  CAS  Google Scholar 

  • Ghannoum MA (2000) Potential role of phospholipases in virulence and fungal pathogenesis. Clin Microbiol Rev 13:122–143

    Article  PubMed  CAS  Google Scholar 

  • Golan DE, Brown CS, Cianci CM, Furlong ST, Caulfied JP (1986) Schistosomula of Schistosoma mansoni use lysophosphatidylcholine to lyse adherent human red blood cells and immobilize red cell membrane components. J Cell Biol 103:819–828

    Article  PubMed  CAS  Google Scholar 

  • Gross RW, Sobel BE (1983) Rabbit myocardial cytosolic lysophospholipase. Purification, characterization, and competitive inhibition by l-palmitoyl carnitine. J Biol Chem 258:5221–5226

    PubMed  CAS  Google Scholar 

  • Lee JS, Kim IS, Sohn WM, Lee J, Yong TS (2006) Vaccination with DNA encoding cysteine proteinase confers protective immune response to rats infected with Clonorchis sinensis. Vaccine 24:2358–2366

    Article  PubMed  CAS  Google Scholar 

  • Li SY, Chung BS, Choi MH, Hong ST (2004) Organ-specific antigens of Clonorchis sinensis. Korean J Parasitol 42:169–174

    Article  PubMed  Google Scholar 

  • Li Y, Zhang YH, Wang S, An XQ (2003) Assay for phospholipase A (PLA2) activity in microemulsions of by AOT water-isooctane. Journal of Nanjing Normal University (Natural Science Ed) 26:44–46

    CAS  Google Scholar 

  • Lun ZR, Gasser RB, Lai DH, Li AX, Zhu XQ, Yu XB, Fang YY (2005) Clonorchiasis: a key foodborne zoonosis in China. Lancet Infect Dis 5:31–41

    Article  PubMed  Google Scholar 

  • Lu T, Ito M, Tchoua U, Takemori H, Okamoto M, Tojo H (2001) Identification of essential residues for catalysis of rat intestinal phospholipase B/lipase. Biochemistry 40:7133–7139

    Article  PubMed  CAS  Google Scholar 

  • Masamune A, Sakai Y, Satoh A, Fujita M, Yoshida M, Shimosegawa T (2001) Lysophosphatidylcholine induces apoptosis in AR42J cells. Pancreas 22:75–83

    Article  PubMed  CAS  Google Scholar 

  • Pickard RT, Chiou XG, Strifler BA, DeFelippis MR, Hyslop PA, Tebbe AL, Yee YK, Reynolds LJ, Dennis EA, Kramer RM, Sharp JD (1996) 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 271:19225–19231

    Article  PubMed  CAS  Google Scholar 

  • Portilla D, Crew MD, Grant D, Serrero G, Bates LM, Dai G, Sasner M, Cheng J, Buonanno A (1998) cDNA cloning and expression of a novel family of enzymes with calcium-independent phospholipase A2 and lysophospholipase activities. J Am Soc Nephrol 9:1178–1186

    PubMed  CAS  Google Scholar 

  • Rao KV, He YX, Kalyanasundaram R (2003) Expression of a 28-kilodalton glutathione S-transferase antigen of Schistosoma mansoni on the surface of filamentous phages and evaluation of its vaccine potential. Clin Diagn Lab Immunol 10:536–541

    Article  PubMed  CAS  Google Scholar 

  • Redmond DL, Knox DP (2006) Further protection studies using recombinant forms of Haemonchus contortus cysteine proteinases. Parasite Immunol 28:213–219

    Article  PubMed  CAS  Google Scholar 

  • Sharp JD, Pickard RT, Chiou XG, Manetta JV, Kovacevic S, Miller JR, Varshavsky AD, Roberts EF, Strifler BA, Brems DN et al (1994) Serine 228 is essential for catalytic activities of 85-kDa cytosolic phospholipase A2. J Biol Chem 269:23250–23254

    PubMed  CAS  Google Scholar 

  • Sim S, Park GM, Yong TS (2003) Cloning and characterization of Clonorchis sinensis myoglobin using immune sera against excretory–secretory antigens. Parasitol Res 91:338–343

    Article  PubMed  Google Scholar 

  • Sunaga H, Sugimoto H, Nagamachi Y, Yamashita S (1995) Purification and properties of lysophospholipase isoenzymes from pig gastric mucosa. Biochem J 308:551–557

    PubMed  CAS  Google Scholar 

  • Wang A, Dennis EA (1999) Mammalian lysophospholipases. Biochim Biophys Acta 1439:1–16

    PubMed  CAS  Google Scholar 

  • Wang A, Deems RA, Dennis EA (1997a) Cloning, expression, and catalytic mechanism of murine lysophospholipase I. J Biol Chem 272:12723–12729

    Article  PubMed  CAS  Google Scholar 

  • Wang A, Loo R, Chen Z, Dennis EA (1997b) Regiospecificity and catalytic triad of lysophospholipase I. J Biol Chem 272:22030–22036

    Article  PubMed  CAS  Google Scholar 

  • Wright LC, Payne J, Santangelo RT, Simpanya MF, Chen SC, Widmer F, Sorrell TC (2004) Cryptococcal phospholipases: a novel lysophospholipase discovered in the pathogenic fungus Cryptococcus gattii. Biochem J 384:377–384

    Article  PubMed  CAS  Google Scholar 

  • Yang G, Yu XB, Wu ZD, Xu J, Song LX, Zhang HM, Hu XC, Zheng NC, Guo LC, Xu J, Dai JF, Ji CN, Gu SH, Ying K (2005) Molecular cloning and characterization of a novel adenylate kinase 3 gene from Clonorchis sinensis. Parasitol Res 95:406–412

    Article  PubMed  Google Scholar 

  • Yamasaki H, Araki K, Lim PK, Zasmy N, Mak JW, Taib R, Aoki T (2000) Development of a highly specific recombinant Toxocara canis second-stage larva excretory–secretory antigen for immunodiagnosis of human toxocariasis. J Clin Microbiol 38:1409–1413

    PubMed  CAS  Google Scholar 

  • Zhang YY, Dennis EA (1988) Purification and characterization of a lysophospholipase from a macrophage-like cell line P388D1. J Biol Chem 263:9965–9972

    PubMed  CAS  Google Scholar 

  • Zheng BY, Feng YY, Xu JC, Zhang YH, An XQ (2005a) The effect of different factors on the specific activity of PLA2 in vesicle solution. Journal of Nanjing Normal University (Natural Science Ed) 28:77–79

    CAS  Google Scholar 

  • Zheng N, Xu J, Wu Z, Chen J, Hu X, Song L, Yang G, Ji C, Chen S, Gu S, Ying K, Yu X (2005b) Clonorchis sinensis: molecular cloning and functional expression of novel cytosolic malate dehydrogenase. Exp Parasitol 109:220–227

    Article  PubMed  CAS  Google Scholar 

  • Zhou L, Shi M, Guo Z, Brisbon W, Hoover R, Yang H (2006) Different cytotoxic injuries induced by lysophosphatidylcholine and 7-ketocholesterol in mouse endothelial cells. Endothelium 13:213–226

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by a grant from the key science and technique program of Guangdong province, China (No. 2004A30801004).

Author information

Authors and Affiliations

  1. Department of Parasitology, Zhongshan School of Medicine, SunYat-sen University, Guangzhou, 510080, People’s Republic of China

    Changling Ma, Xuchu Hu, Fengyu Hu, Yanwen Li, Xiaoxiang Chen, Zhenwen Zhou, Fangli Lu, Jin Xu, Zhongdao Wu & Xinbing Yu

  2. Department of Microbiology and Parasitology, Guangzhou Medical College, Guangzhou, 510182, People’s Republic of China

    Changling Ma & Xiaoxiang Chen

Authors
  1. Changling Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xuchu Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Fengyu Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yanwen Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Xiaoxiang Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Zhenwen Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Fangli Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jin Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Zhongdao Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Xinbing Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Xinbing Yu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ma, C., Hu, X., Hu, F. et al. Molecular characterization and serodiagnosis analysis of a novel lysophospholipase from Clonorchis sinensis . Parasitol Res 101, 419–425 (2007). https://doi.org/10.1007/s00436-007-0481-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00436-007-0481-3

Keywords

Search

Navigation