Animal cells and systems

  • 제6권4호
  • /
  • Pages.341-347
  • /
  • 2002
  • /
  • 1976-8354(pISSN)
  • /
  • 2151-2485(eISSN)
한국통합생물학회 (The Korean Society for Integrative Biology)
권호 표지

Changes in Profiles of Major Proteins in Encysting Acanthamoeba castellanii

  • Park, Joon-Tae (School of Biological Sciences, Seoul National University) ;
  • Jeong, Young-Eui (School of Biological Sciences, Seoul National University) ;
  • Ahn, Tae-In (School of Biological Sciences, Seoul National University)
  • 발행 : 2002.12.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

The life cycle of Acanthamoeba is comprised of two distinct stages, tropho-zoite and cyst. During periods of stress, trophozoites undergo cellular differentiation into cyst. In order to understand the cellular differentiation, ore followed changes in profiles of major proteins by 2D-PAGE and ubiqui-tinated proteins by immunoblotting with anti-ubiquitin (Ub) monoclonal antibody (mAb) as a probe. We observed 51 proteins present in trophozoite were lost with the encystment. We found that 43 proteins within 24 h, and 8 proteins in 96 h of encystment. Among them, 17 proteins were staine with anti-Ub mAb. In cysts, 16 proteins including 2 anti-Ub mAb-reactive proteins were newly synthesized. Four proteins were newly detected in 24 h-cyst and disappeared in 96 h-cyst, one protein was synthesized in 24-96 h-cyst and disappeared in 168 h-cyst, and 11 proteins appeared upon en-cystment and were present in all cyst stages. We identified a cyst specific 33 kDa protein as subtilisin-like serine proteinase by N-terminal sequencing. Identification of these proteins lost and newly synthesized with encystment would improve our understanding of cysting protozoan parasites.

키워드

참고문헌

  1. Bowers B and Korn ED (1969) The fine structure of Acanthamoeba castellanii (Neff strain) II encystment. J Cell Biol 41: 786-805 https://doi.org/10.1083/jcb.41.3.786
  2. Corliss JO and Esser SC (1984) Comments on the role of the cysts in the life cycle and survival of free-living protozoa. Trans Am Microsc Soc 93: 578-593 https://doi.org/10.2307/3225158
  3. Freshney RI (1994) Culture of Animal Cells. Wiley-Liss, New York
  4. Griffiths AJ and Hughes DE (1969) The physiology of encystment of Hartmannella castellanii. J Protozool 16: 93-99 https://doi.org/10.1111/j.1550-7408.1969.tb02238.x
  5. Gonzalez J, Bai G, Frevert U, Corey EJ, and Eichinger D (1999) Proteasome-dependent cyst formation and stagespecific Ub mRNA accumulation in Entamoeba invadens. Eur J Biochem 264: 897-904 https://doi.org/10.1046/j.1432-1327.1999.00682.x
  6. Hirukawa Y, Nakato H, Izumi S, Tsuruhara T, and Tomino S (1998) Structure and expression of a cyst specific protein of Acanthamoeba castellanii. Biochim Biophys Acta 1398: 47-56 https://doi.org/10.1016/S0167-4781(98)00026-8
  7. Hong YC, Kong HH, Ock MS, Kim IS, and Chung DI (2000) Isolation and characterization of a cDNA encoding a subtilisinlike serine proteinase (ahSUB) from Acanthamoeba healyi. Mol Biochem Parasitol 111: 441-446 https://doi.org/10.1016/S0166-6851(00)00326-1
  8. Jantzen H (1981) Control of actin synthesis during the development of Acanthamoeba castellanii. Dev Biol 82: 113-126 https://doi.org/10.1016/0012-1606(81)90433-4
  9. Jensen TJ, Loo MA, Pind S, Williams DB, Goldberg AL, and Riordan JR (1995) Multiple proteolytic systems, including the proteasome, contribute to CFTR processing. Cell 83: 129-135 https://doi.org/10.1016/0092-8674(95)90241-4
  10. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head bacteriophage T4. Nature 227: 680-685 https://doi.org/10.1038/227680a0
  11. Marciano-Cabral F, Puffenbarger R, and Cabral GA (2000) The increasing importance of Acanthamoeba infections. J Euk Microbial 47: 29-36 https://doi.org/10.1111/j.1550-7408.2000.tb00007.x
  12. Martieze AJ (1985) Free-living amoebas. In: Ration B (ed), Natural History, Prevention, Diagnosis, Pathology and Treatment of Disease, CRC Press, pp 5-42
  13. Neff RJ, Ray SA, Benton WF, and Wilborn M (1964) Induction of synchronous encystment (differentiation) in Acanthamoeba sp. In: Prescott DM (ed), Methods in Cell Physiology I, Academic Press Inc, New York, pp 55
  14. Neff RJ and Neff RH (1969) The biochemistry of amoebic encystment. Symp Soc Exp Biol 23: 51-81
  15. O'Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250: 4007-4021
  16. Parsell DA and Lindquist S (1993) The function of heat-shock proteins in stress tolerance: degradation and reactivation of damaged proteins. Ann Rev Genet 27: 437-496 https://doi.org/10.1146/annurev.ge.27.120193.002253
  17. Ramagli LS and Rodriguez LV (1985) Quantitation of microgram amounts of protein in two dimensional polyacrylamide gel electrophoresis sample buffer. Electrophoresis 6: 559-563 https://doi.org/10.1002/elps.1150061109
  18. Selgen PO and Bohley P (1992) Autophagy and other vacuolar protein degradation mechanisms. Experientia 48: 158-172 https://doi.org/10.1007/BF01923509
  19. Stratford MP and Griffiths AJ (1978) Variations in the properties and morphology of cysts of Acanthamoeba castellanii. J Gen Microbial 108: 33-37 https://doi.org/10.1099/00221287-108-1-33
  20. Weisman RA (1976) Differentiation in Acanthamoeba castellanii. Ann Rev Microbial 30: 189-219 https://doi.org/10.1146/annurev.mi.30.100176.001201