PURIFICATION AND ACTIVITY ASSAY OF PROTEIN DISULPHIDE-ISOMERASE

Hu, Chao-hong Wang, Zhi-zhen

Chinese Journal of Biochemistry and Molecular Biology ›› 1988, Vol. 4 ›› Issue (01) : 61-67.

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Chinese Journal of Biochemistry and Molecular Biology ›› 1988, Vol. 4 ›› Issue (01) : 61-67.
Article

PURIFICATION AND ACTIVITY ASSAY OF PROTEIN DISULPHIDE-ISOMERASE

  • Hu, Chao-hong Wang, Zhi-zhen
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Abstract

Protein disulphide-isomerase(PDI, EC 5.3.4.1) has been isolated and purified from bovine liver with a yield of 98 mg/500g fresh tissue. The enzyme is homogeneous and has Mr 62,000 as determined by sodium dodecyl sulfate/polyacrylamide gel electrophoresis in the presence of dithiothreitol. The molar extinction coefficient of the enzyme has been found at 280nm and pH 7.5 to be 1.03×105, An activity assay method is employed in which the reactivation of "scrambled" ribonu-clease A is monitored as the increase of absorbance at 260nm due to the hydrolysis of yeast RNA catalysed by reactivated ribonuclease A.This method is simple and more sensitive than monitoring the increase in absorbance at 260nm relative to 280nm with a dual-wavelength instrument.

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Protein disulphide-isomerase

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Hu, Chao-hong Wang, Zhi-zhen. PURIFICATION AND ACTIVITY ASSAY OF PROTEIN DISULPHIDE-ISOMERASE[J]. Chinese Journal of Biochemistry and Molecular Biology, 1988, 4(01): 61-67

References

[1] Golderger, R. F. Epstein, C. J. and Anfinsen, C. B.(1963) , J. Biol. Chem., 239, 628-635. [2] Venetianer, P. & Straub, F. B. (1963) , Biochim. Biophys. Acta, 67, 166-168. [3] Tomizawa, H.H(1962) , J. Biol. Chem., 237, 428-431. [4] Lambert, N. and Freedman, R. B. (1983) , Biochem. J., 213, 235-243. [5] Bjelland, S., Wallevik, K., Kroll, J., Dixon, J.E., Morin, J.E., Freedman, R. B., Lambert, N., Varndani, P.T., Nafz, M.A.(1983) , Biochimica et Biophysics Acta, 747,197-199. [6] Freedman,R.B.,Brockway, B. E.and Lambert, N. (1984) , Biochemical Society Transactions, 12, 929-932. [7] Freedman, R.B. (1994) , TIBS, 438-441. [8] Lambert, N.and Freedman, R.B. (1983) , Biochem. J., 213, 225-234. [9] Mills, E. N. C., Lambert, N. and Freedmao, R.B.(1983) , Biochem. J., 213, 244-248. [10] Edman, J. C., Ellis, L., Blacher, R.W., Roth, R. A.and Rutter, W. J.(1985) , Nature, 317, 267-270. [11] Morangos, P. J. and Constantinides, S. M (1974) , Biochemistry, 13, 904-910. [12] Anfinsen, C. B. and Haber, E. (1961) , J. Biol.Chem., 236, 1361-1363. [13] Haber, E. and Anfinsen, C.B. (1962) , J.Biol.Chem., 237, 1839-1844. [14] Ellman, G.L. (1959) , Aach. Biochem. Biophys. 82, 70-77. [15] Ibbetson, A.L.and Freedman, R.B. (1976) , Biochem. J., 159, 377-384. [16] Lowry, O.H., Roseborough, N.J., Farr, A. L.and Randall, R.J. (1951) , J. Biol. Chem.,193, 265-275. [17] an Hdbook of Biochemistry, Selected Data for Molecular Biology second edition, Edited by Sober, H. A., p.C-71. [18] 徐秀璋(1983) ,《分析测试通报》.第2卷,第3期.8-14页. [19] Penke, B. (1974) , Anal. Biochem. 60, 45-50. [20] Inglis, A.S.and Liu, T.Y. (1970) , J. Biol. Chem., 245, 112-116. [21] Hillson, D.A., Lambert, N. and Freedman, R.B. (1984) , Methods in Enzymology, 107, 281-294.
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