Parasites, Hosts and Diseases

The Korea Society for Parasitology and Tropical Medicine (대한기생충학·열대의학회)
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Cloning and characterization of Giardia intestinalis cyclophilin

  • Yu, Hak-Sun (Department of Parasitology, College of Medicine, Kyungpook national University) ;
  • Kong, Hyun-Hee (Department of Parasitology, College of Medicine, Kyungpook national University) ;
  • Chung, Dong-Il (Department of Parasitology, College of Medicine, Kyungpook national University)
  • Published : 2002.09.01
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Abstract

The cyclophilins (Cyps) are family members of proteins that exhibit peptidylprolyl cis-trans isomerase (PPIase, EC 5.2.1.8) activity and bind the immunosuppressive agent cyclosprin A (CsA) in varying degrees. During the process of random sequencing of a cDNA library made from Giardia intestinalis WB strain, the cyclophilin gene (gicypl) was isolated. An open reading frame of gicyp1 gene was 576 nucleotides, which corresponded to a translation product of 176 amino acids (Gicypl). The identity with other Cyps was about 58-71%. The 13 residues that constituted the CsA binding site of human cyclophilin were also detected in the amino acid sequence of Gicypl, including tryptophan residue essential for the drug binding. The single copy of the gicypl gene was detected in the G. intestinalis chromosome by southern hybridization analysis. Recombinant Gicyp 1 protein clearly accelerated the rate of cis ${\rightarrow}$ trans isomerization of the peptide substrate and the catalysis was completely inhibited by the addition of $0.5{\;}{\mu}M$ CsA.

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References

  1. Adam RD (2001) Biology of Giardia lamblia. Clin Microbiol Rev 14: 447-475
  2. Bergsma DJ, Eder C, Gross M, et al. (1991) The cyclophilin multigene family of peptidylprolyl isomerase. J Biol Chem 266: 23204- 23214
  3. Chappell LH, Thomson AW (1988) Studies on the action of cyclosporin A against Schstosoma mansoni and other parasitic infections. Transplant Proc 20: 291-297
  4. Clipstone NA, Crabtree GR (1992) Identification of calcineurin as a key signaling enzyme in Tlymphocyte activation. Nature 357: 695-697
  5. Duina AA, Marsh JA, Kurtz RB, Chang HJ, Lindquist S, Gaber RF (1998) The peptidylprolyl isomerase domain of the CyP-40 cyclophilin homology Cpr7 is not required to support growth or glucocorticoid receptor activity in Saccharomyces cerevisiae. J Biol Chem 273: 10819-10822
  6. Fischer G, Wittmann-Liebold B, Lang K, Keifhaber T, Schmid FX (1989) Cyclophilin and peptidyl-prolyl cis trans isomerase are probably identical proteins. Nature 337: 476- 478
  7. Gasser CS, Gunning DA, Budelier KA, Brown SM (1990) Structure and expression of cytosolic cyclophilin/peptidyl-prolyl cis-trans isomerase of higher plants and production of active tomato cyclophilin in Escherichia coli. Proc Natl Acad Sci USA. 87: 9519-9523
  8. Haendler B, Hofer-Warbinek R, Hofer E (1987) Complementary DNA for human T-cell cyclophilin. EMBO J 6: 947-950
  9. Handschumacher RE, Harding MW, Rice J, Drugge RJ, Speicher DW (1984) Cyclophilin: a specific cytosolic binding protein for cyclosporin A. Science 226: 544-547
  10. Kocken CH, van der Wel A, Rosenwirth B, Thomas AW (1996) Plasmodium vivax: in vitro antiparasitic effect of cyclosporins. Exp Parasitol 84: 439-443
  11. Liu J, Walsh CT (1990) Peptidyl-prolyl cis-transisomerase from Escherichia coli: a periplasmic homolog of cyclophilin that is not inhibited by cyclosporin A. Proc Natl Acad Sci USA 87: 4028-4032
  12. Liu J, Chen CM, Walsh CT (1991) Human and Escherichia coli cyclophilins: sensitivity to inhibition by the immunosuppressant cyclosporinA correlates with a specific tryptophan residue. Biochemistry 30: 2306- 2310 https://doi.org/10.1021/bi00223a003
  13. Minotto L, Tutticci EA, Bagnara AS, Schofield PJ, Edwards MR (1999) Characterization and expression of the carbamate kinase gene from Giardia intestinalis. Mol Biochem Parasitol 98: 43-51
  14. Ostoa-Saloma P, Cesar Carrero J, Petrossian P, Herion P, Landa A, Pedro Laclette J (2000) Cloning, characterization and functional expression of a cyclophilin of Entamoeba histolytica. Mol Biochem Parasitol 107: 219- 225
  15. Page AP, Kumar S, Carlow CKS (1995) Parasite cyclophilins and antiparasite activity of cyclosporin A. Parasitol Today 11: 385-389
  16. Reddy GR (1995) Cloning and characterization of a Plasmodium falciparum cyclophilin gene that is stage-specifically expressed. Mol Biochem Parasitol 73: 111-121
  17. Schneider H, Charara N, Schmitz R, et al. (1994) Human cyclophilin C: Primary structure tissue distribution and determination of binding specificity for cyclosporins. Biochemistry 33: 8218-8224
  18. Smith DB, Johnson KS (1988) Single-step purification of polypeptides expressed in Escherichia coli as fusion proteins with glutathione-S-transferase. Gene 67: 31-40
  19. Sogin ML, Hinkle G, Leipe DD (1993) Universal tree of life. Nature 362: 795
  20. Solbach W, Forberg K, Rollinghoff M (1986) Effect of T-lymphocyte suppression on the parasite burden in Leishmania major-infected, genetically susceptible BALB/c mice. Infect Immun 54: 909-912
  21. Spik G, Haendler B, Delmas O, et al. (1991) A novel secreted cyclophilin-like protein (SCYLP). J Biol Chem 266: 10735-10738
  22. Trandinh CC, Pao GM, Saier MH Jr (1992) Structural and evolutionary relationships among the immunophilins: two ubiquitous families of peptidyl-prolyl cis-trans isomerases. FASEB J 6: 3410-3420
  23. Yee J, Mowatt MR, Dennis PP, Nash TE (2000) Transcriptional analysis of the glutamate dehydrogenase gene in the primitive eukaryote, Giardia lamblia. Identification of a primordial gene promoter. J Biol Chem 275: 11432-11439