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Molecular and Biochemical Characterization of Opisthorchis viverrini Calreticulin

  • Chaibangyang, Wanlapa (Graduate Program in Biomedical Sciences, Faculty of Allied Health Sciences, Thammasat University) ;
  • Geadkaew-Krenc, Amornrat (Graduate Program in Biomedical Sciences, Faculty of Allied Health Sciences, Thammasat University) ;
  • Vichasri-Grams, Suksiri (Department of Biology, Faculty of Science, Mahidol University) ;
  • Tesana, Smarn (Food-borne Parasite Research Group, Department of Parasitology, Faculty of Medicine, Khon Kaen University) ;
  • Grams, Rudi (Graduate Program in Biomedical Sciences, Faculty of Allied Health Sciences, Thammasat University)
  • 투고 : 2017.11.13
  • 심사 : 2017.12.18
  • 발행 : 2017.12.31

초록

Calreticulin (CALR), a multifunctional protein thoroughly researched in mammals, comprises N-, P-, and C-domain and has roles in calcium homeostasis, chaperoning, clearance of apoptotic cells, cell adhesion, and also angiogenesis. In this study, the spatial and temporal expression patterns of the Opisthorchis viverrini CALR gene were analyzed, and calcium-binding and chaperoning properties of recombinant O. viverrini CALR (OvCALR) investigated. OvCALR mRNA was detected from the newly excysted juvenile to the mature parasite by RT-PCR while specific antibodies showed a wide distribution of the protein. OvCALR was localized in tegumental cell bodies, testes, ovary, eggs, Mehlis' gland, prostate gland, and vitelline cells of the mature parasite. Recombinant OvCALR showed an in vitro suppressive effect on the thermal aggregation of citrate synthase. The recombinant OvCALR C-domain showed a mobility shift in native gel electrophoresis in the presence of calcium. The results imply that OvCALR has comparable function to the mammalian homolog as a calcium-binding molecular chaperone. Inferred from the observed strong immunostaining of the reproductive tissues, OvCALR should be important for reproduction and might be an interesting target to disrupt parasite fecundity. Transacetylase activity of OvCALR as reported for calreticulin of Haemonchus contortus could not be observed.

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참고문헌

  1. Fliegel L, Burns K, MacLennan DH, Reithmeier RA, Michalak M. Molecular cloning of the high affinity calcium-binding protein (calreticulin) of skeletal muscle sarcoplasmic reticulum. J Biol Chem 1989; 264: 21522-21528.
  2. Smith MJ, Koch GL. Multiple zones in the sequence of calreticulin (CRP55, calregulin, HACBP), a major calcium binding ER/SR protein. EMBO J 1989; 8: 3581-3586.
  3. Munro S, Pelham HR. A C-terminal signal prevents secretion of luminal ER proteins. Cell 1987; 48: 899-907. https://doi.org/10.1016/0092-8674(87)90086-9
  4. Michalak M, Groenendyk J, Szabo E, Gold LI, Opas M. Calreticulin, a multi-process calcium-buffering chaperone of the endoplasmic reticulum. Biochem J 2009; 417: 651-666. https://doi.org/10.1042/BJ20081847
  5. Raghavan M, Wijeyesakere SJ, Peters LR, Del Cid N. Calreticulin in the immune system: ins and outs. Trends Immunol 2013; 34: 13-21. https://doi.org/10.1016/j.it.2012.08.002
  6. Lu YC, Weng WC, Lee H. Functional roles of calreticulin in cancer biology. Biomed Res Int 2015; 2015: 526524.
  7. de Bruyn M, Wiersma VR, Helfrich W, Eggleton P, Bremer E. The ever-expanding immunomodulatory role of calreticulin in cancer immunity. Front Oncol 2015; 5: 35.
  8. Khalife J, Pierce RJ, Godin C, Capron A. Cloning and sequencing of the gene encoding Schistosoma mansoni calreticulin. Mol Biochem Parasitol 1993; 62: 313-315. https://doi.org/10.1016/0166-6851(93)90120-M
  9. Khalife J, Liu JL, Pierce R, Porchet E, Godin C, Capron A. Characterization and localization of Schistosoma mansoni calreticulin Sm58. Parasitology 1994; 108: 527-532. https://doi.org/10.1017/S0031182000077398
  10. Hooker CW, Brindley PJ. Cloning of a cDNA encoding SjIrV1, a Schistosoma japonicum calcium-binding protein similar to calnexin, and expression of the recombinant protein in Escherichia coli. Biochim Biophys Acta 1999; 1429: 331-341. https://doi.org/10.1016/S0167-4838(98)00233-7
  11. Scott JC, McManus DP. Molecular cloning and functional expression of a cDNA encoding the major endoplasmic reticulumassociated calcium-binding protein, calreticulin, from Philippine strain Schistosoma japonicum. Parasitol Int 1999; 48: 35-46. https://doi.org/10.1016/S1383-5769(98)00039-7
  12. Singh P, Ponnan P, Krishnan S, Tyagi TK, Priya N, Bansal S, Scumaci D, Gaspari M, Cuda G, Joshi P, Gambhir JK, Saluja D, Prasad AK, Saso L, Rastogi RC, Parmar VS, Raj HG. Protein acyltransferase function of purified calreticulin. Part 1: characterization of propionylation of protein utilizing propoxycoumarin as the propionyl group donor. J Biochem 2010; 147: 625-632. https://doi.org/10.1093/jb/mvq002
  13. Singh P, Ponnan P, Priya N, Tyagi TK, Gaspari M, Krishnan S, Cuda G, Joshi P, Gambhir JK, Sharma SK, Prasad AK, Saso L, Rastogi RC, Parmar VS, Raj HG. Protein acyltransferase function of purified calreticulin: the exclusive role of P-domain in mediating protein acylation utilizing acyloxycoumarins and acetyl CoA as the acyl group donors. Protein Pept Lett 2011; 18: 507-517. https://doi.org/10.2174/092986611794927938
  14. Young ND, Campbell BE, Hall RS, Jex AR, Cantacessi C, Laha T, Sohn WM, Sripa B, Loukas A, Brindley PJ, Gasser RB. Unlocking the transcriptomes of two carcinogenic parasites, Clonorchis sinensis and Opisthorchis viverrini. PLoS Negl Trop Dis 2010; 4: e719. https://doi.org/10.1371/journal.pntd.0000719
  15. Rice P, Longden I, Bleasby A. EMBOSS: the European Molecular Biology Open Software Suite. Trends Genet 2000; 16: 276-277. https://doi.org/10.1016/S0168-9525(00)02024-2
  16. Sievers F, Wilm A, Dineen D, Gibson TJ, Karplus K, Li W, Lopez R, McWilliam H, Remmert M, Soding J, Thompson JD, Higgins DG. Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Mol Syst Biol 2011; 7: 539.
  17. Petersen TN, Brunak S, von Heijne G, Nielsen H. SignalP 4.0: discriminating signal peptides from transmembrane regions. Nat Methods 2011; 8: 785-786. https://doi.org/10.1038/nmeth.1701
  18. Sigrist CJ, de Castro E, Cerutti L, Cuche BA, Hulo N, Bridge A, Bougueleret L, Xenarios I. New and continuing developments at PROSITE. Nucleic Acids Res 2013; 41: 344-347.
  19. UniProt Consortium. UniProt: a hub for protein information. Nucleic Acids Res 2015; 43: 204-212. https://doi.org/10.1093/nar/gku989
  20. Saito Y, Ihara Y, Leach MR, Cohen-Doyle MF, Williams DB. Calreticulin functions in vitro as a molecular chaperone for both glycosylated and non-glycosylated proteins. EMBO J 1999; 18: 6718-6729. https://doi.org/10.1093/emboj/18.23.6718
  21. Vichasri-Grams S, Subpipattana P, Sobhon P, Viyanant V, Grams R. An analysis of the calcium-binding protein 1 of Fasciola gigantica with a comparison to its homologs in the phylum Platyhelminthes. Mol Biochem Parasitol 2006; 146: 10-23. https://doi.org/10.1016/j.molbiopara.2005.10.012
  22. Bansal S, Ponnan P, Raj HG, Weintraub ST, Chopra M, Kumari R, Saluja D, Kumar A, Tyagi TK, Singh P, Prasad AK, Saso L, Rastogi RC, Parmar VS. Autoacetylation of purified calreticulin transacetylase utilizing acetoxycoumarin as the acetyl group donor. Appl Biochem Biotechnol 2009; 157: 285-298. https://doi.org/10.1007/s12010-008-8357-2
  23. Kathuria A, Gupta A, Priya N, Singh P, Raj HG, Prasad AK, Parmar VS, Sharma SK. Specificities of calreticulin transacetylase to acetoxy derivatives of 3-alkyl-4-methylcoumarins: effect on the activation of nitric oxide synthase. Bioorg Med Chem 2009; 17: 1550-1556. https://doi.org/10.1016/j.bmc.2009.01.003
  24. Young ND, Nagarajan N, Lin SJ, Korhonen PK, Jex AR, Hall RS, Safavi-Hemami H, Kaewkong W, Bertrand D, Gao S, Seet Q, Wongkham S, Teh BT, Wongkham C, Intapan PM, Maleewong W, Yang X, Hu M, Wang Z, Hofmann A, Sternberg PW, Tan P, Wang J, Gasser RB. The Opisthorchis viverrini genome provides insights into life in the bile duct. Nat Commun 2014; 5: 4378. https://doi.org/10.1038/ncomms5378
  25. Baksh S, Michalak M. Expression of calreticulin in Escherichia coli and identification of its Ca2+ binding domains. J Biol Chem 1991; 266: 21458-21465.
  26. Park BJ, Lee DG, Yu JR, Jung SK, Choi K, Lee J, Lee J, Kim YS, Lee JI, Kwon JY, Lee J, Singson A, Song WK, Eom SH, Park CS, Kim DH, Bandyopadhyay J, Ahnn J. Calreticulin, a calcium-binding molecular chaperone, is required for stress response and fertility in Caenorhabditis elegans. Mol Biol Cell 2001; 12: 2835-2845. https://doi.org/10.1091/mbc.12.9.2835
  27. Leach MR, Cohen-Doyle MF, Thomas DY, Williams DB. Localization of the lectin, ERp57 binding, and polypeptide binding sites of calnexin and calreticulin. J Biol Chem 2002; 277: 29686-29697. https://doi.org/10.1074/jbc.M202405200
  28. Raj HG, Kohli E, Goswami R, Goel S, Rastogi RC, Jain SC, Wengel J, Olsen CE, Parmar VS. Mechanism of biochemical action of substituted benzopyran-2-ones. Part 8: Acetoxycoumarin: protein transacetylase specificity for aromatic nuclear acetoxy groups in proximity to the oxygen heteroatom. Bioorg Med Chem 2001; 9: 1085-1089. https://doi.org/10.1016/S0968-0896(00)00328-X
  29. Kozlov G, Pocanschi CL, Rosenauer A, Bastos-Aristizabal S, Gorelik A, Williams DB, Gehring K. Structural basis of carbohydrate recognition by calreticulin. J Biol Chem 2010; 285: 38612-38620. https://doi.org/10.1074/jbc.M110.168294
  30. Chouquet A, Paidassi H, Ling WL, Frachet P, Houen G, Arlaud GJ, Gaboriaud C. X-ray structure of the human calreticulin globular domain reveals a peptide-binding area and suggests a multimolecular mechanism. PLoS One 2011; 6: e17886. https://doi.org/10.1371/journal.pone.0017886
  31. Schrag JD, Bergeron JJ, Li Y, Borisova S, Hahn M, Thomas DY, Cygler M. The Structure of calnexin, an ER chaperone involved in quality control of protein folding. Mol Cell 2001; 8: 633-644. https://doi.org/10.1016/S1097-2765(01)00318-5
  32. Kumari R, Bansal S, Gupta G, Arora S, Kumar A, Goel S, Singh P, Ponnan P, Priya N, Tyagi TK, Baghel AS, Manral S, Tandon R, Joshi R, Rohil V, Gaspari M, Kohli E, Tyagi YK, Dwarakanath BS, Saluja D, Chatterji S, Sharma SK, Prasad AK, Rastogi RC, Raj HG, Parmar VS. Calreticulin transacylase: genesis, mechanism of action and biological applications. Biochimie 2010; 92: 1173-1179. https://doi.org/10.1016/j.biochi.2010.01.016
  33. Apinhasmit W, Sobhon P, Saitongdee P, Menayotin S, Upatham ES. Opisthorchis viverrini: ultrastructure of the tegument of the first-week juveniles and adult flukes. Int J Parasitol 1994; 24: 613-621. https://doi.org/10.1016/0020-7519(94)90113-9
  34. Kaewpitoon N, Laha T, Kaewkes S, Yongvanit P, Brindley PJ, Loukas A, Sripa B. Characterization of cysteine proteases from the carcinogenic liver fluke, Opisthorchis viverrini. Parasitol Res 2008; 102: 757-764. https://doi.org/10.1007/s00436-007-0831-1
  35. Sripa J, Laha T, To J, Brindley PJ, Sripa B, Kaewkes S, Dalton JP, Robinson MW. Secreted cysteine proteases of the carcinogenic liver fluke, Opisthorchis viverrini: regulation of cathepsin F activation by autocatalysis and trans-processing by cathepsin B. Cell Microbiol 2010; 12: 781-795. https://doi.org/10.1111/j.1462-5822.2010.01433.x
  36. Mesaeli N, Nakamura K, Zvaritch E, Dickie P, Dziak E, Krause KH, Opas M, MacLennan DH, Michalak M. Calreticulin is essential for cardiac development. J Cell Biol 1999; 144: 857-868. https://doi.org/10.1083/jcb.144.5.857
  37. Guo L, Nakamura K, Lynch J, Opas M, Olson EN, Agellon LB, Michalak M. Cardiac-specific expression of calcineurin reverses embryonic lethality in calreticulin-deficient mouse. J Biol Chem 2002; 277: 50776-50779. https://doi.org/10.1074/jbc.M209900200
  38. Lee W, Kim KR, Singaravelu G, Park BJ, Kim DH, Ahnn J, Yoo YJ. Alternative chaperone machinery may compensate for calreticulin/calnexin deficiency in Caenorhabditis elegans. Proteomics 2006; 6: 1329-1339. https://doi.org/10.1002/pmic.200500320
  39. Frickel EM, Riek R, Jelesarov I, Helenius A, Wuthrich K, Ellgaard L. TROSY-NMR reveals interaction between ERp57 and the tip of the calreticulin P-domain. Proc Natl Acad Sci USA 2002; 99: 1954-1959. https://doi.org/10.1073/pnas.042699099

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