플로래스카민과 알파이미저를 이용한 PrP 106-126 펩타이드 정량에 관한 연구

정근홍; 정우영; 계영식; Keunhong Jeong; Woo Young Chung; Young Sik Kye

Journal of the Korean Industrial and Engineering Chemistry, Vol.20, No.6, 628-631, December, 2009

Export Citation

플로래스카민과 알파이미저를 이용한 PrP 106-126 펩타이드 정량에 관한 연구

A Study on the Quantification of PrP 106-126 Peptide by Fluorescamine and Alpha-imager

정근홍^†, 정우영, 계영식

육군사관학교

Keunhong Jeong^†, Woo Young Chung, and Young Sik Kye

Department of Chemistry, Korea Military Academy, Seoul 139-799, Korea

E-mail:

초록

최근 광우병 파동과 프리온 단백질의 전염으로 인하여 발생되는 것으로 알려진 크로이츠펠트 야콥병에 대하여 폭넓게 연구되면서 생화학계에서 프리온 단백질에 대한 관심은 상당하다. 본 연구에서는 프리온 단백질의 일부분인 PrP 106-126의 마이크로 몰농도 단위의 정량분석을 실시하였다. 본 연구에서 플로래스카민은 일차 아민과 반응하여 형광을 띠는 물질로써, 알파이미저는 형광의 세기를 측정하는 기기로써 사용되었다. 따라서 합성된 PrP 106-126으로 플로래스카민과 알파이미저를 이용하여 마이크로 몰농도단위의 정량 분석을 위한 조건을 정립하였으며 이를 통하여 표준 곡선을 얻을 수 있었다. 이 방법은 차후 변형 프리온 단백질에 대한 응집저해제 및 의약품 연구에 큰 기여를 할 것이다.

Recent social interests on mad cow disease and Creutzfelt-Jacob disease lead researcher’s focus onto pathogenic prion proteins causing those diseases. The purpose of this study is to introduce a novel method for micromolecular level quantification of PrP 106-126 peptide, which is a part of prion protein. Fluorescamine has been chosen due to its fluorescence emission characteristics upon reaction with primary amines and Alpha-imager is used to detect the intensity of fluorescence. We succeeded in setting optimal conditions for quantification of PrP 106-206, amyloidogenic prion peptide, at micromolecular level. This study will contribute to identify prion protein aggregation inhibitor and develop new prion protein drug.

Keywords:PrP 106-126;fluorescamine;alpha-imager

[References]

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Prusiner SB, Proc. Natl. Acad. Sci. U S A., 95, 13363 (1998)
Sikkink LA, Ramirez-Alvarado M, AAPG Bull., 13525 (2008)
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McCaman MW, Robins E, J. Lab. Clin. Med., 59, 885 (1962)
Udenfriend S, Stein S, Bohlen P, Dairman W, Leimgruber W, Weigele M, Science, 178, 871 (1971)
Suh J, Chei WS, Lee T, Kim MG, Yoo SH, Jeong K, Ahn JY, J. Biol. Inorg. Chem., 13, 693 (2008)
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Jeong K, Kim MK, Chung WY, Kye YS, Kim JH, Suh J, J. Ind. Eng. Chem., 15(3), 342 (2009)

[Referenced By]

[1] Blackstock JC, Biochemistry, Butterworth-Heinemann, Boston (1998)

[2] Bradford M, Anal. Biochem., 72, 248 (1976)

[3] Lowry OH, Rosebrough NJ, Farr AL, Randall RJ, J. Biol. Chem., 193, 265 (1951)

[4] Hartree EE, Anal. Biochem., 48, 422 (1972)

[5] Wilson K, Walker J, Practical Biochemistry: Principles and Techniques, Cambridge University Press, Cambridge (2000)

[6] Stoscheck CM, Meth. Enz., 182, 50 (1990)

[7] Slater RJ, Experiments in Molecular Biology, Humana Press, New Jersey (1986)

[8] Deriziotis P, Tabrizi SJ, Biochim Biophys. Acta, 1782, 713 (2008)

[9] Wilesmith JW, Wells GA, Cranwell MP, Ryan JB, Vet. Rec., 123, 638 (1988)

[10] Collinge J, J. Neurol. Neurosurg. Psychiatry, 76, 906 (2005)

[11] Wadsworth J, Collinge J, Biochim. Biophys. Acta, 1772, 598 (2007)

[12] Mead S, Eur. J. Hum. Genet., 14, 273 (2006)

[13] Will RG, Ironside JW, Zeidler M, Cousens SN, Estibeiro K, Alperovitch A, Poser S, Pocchiari M, Hofman A, Smith PG, Lancet., 347, 921 (1996)

[14] Collinge J, Sidle KC, Meads J, Ironside J, Hill AF, Nature, 383(6602), 685 (1996)

[15] Hill AF, Desbruslais M, Joiner S, Sidle KC, Gowland I, Collinge J, Doey LJ, Lantos P, Nature, 389(6650), 448 (1997)

[16] Bruce ME, Will RG, Ironside JW, Mcconnell I, Drummond D, Suttie A, Mccardle L, Chree A, Hope J, Birkett C, Cousens S, Fraser H, Bostock CJ, Nature, 389(6650), 498 (1997)

[17] Tagliavini F, Forloni G, D’Ursi P, Bugiani O, Salmona M, Adv. Protein Chem., 57, 171 (2001)

[18] Belosi B, Gaggelli E, Guerrini R, Kozlowski H, Łuczkowski M, Mancini FM, Remelli M, Valensin D, Valensin G, Chem. BioChem., 5, 349 (2004)

[19] Jones CE, Abdelraheim SR, Brown DR, Viles JH, J. Biol. Chem., 279, 32018 (2004)

[20] Prusiner SB, Proc. Natl. Acad. Sci. U S A., 95, 13363 (1998)

[21] Sikkink LA, Ramirez-Alvarado M, AAPG Bull., 13525 (2008)

[22] Suh J, Yoo SH, Kim MG, Jeong K, Ahn JY, Kim MS, Chae PS, Lee TY, Lee J, Lee J, Jang YA, Ko EH, Angew. Chem. Int. Ed., 46, 7064 (2007)

[23] McCaman MW, Robins E, J. Lab. Clin. Med., 59, 885 (1962)

[24] Udenfriend S, Stein S, Bohlen P, Dairman W, Leimgruber W, Weigele M, Science, 178, 871 (1971)

[25] Suh J, Chei WS, Lee T, Kim MG, Yoo SH, Jeong K, Ahn JY, J. Biol. Inorg. Chem., 13, 693 (2008)

[26] Suh J, Lee J, Yoo SH, Jeong K, Lee TY, Ahn JY, Bull. Korean Chem. Soc., 29, 884 (2008)

[27] Jeong K, Kim MK, Chung WY, Kye YS, Kim JH, Suh J, J. Ind. Eng. Chem., 15(3), 342 (2009)