A spectrophotometric assay for superoxide dismutase activities in crude tissue fractions.
作者： H Kuthan , H J Haussmann , J Werringloer
DOI: 10.1042/BJ2370175
关键词:
摘要: A sensitive and reliable assay method was developed to characterize crude cell homogenates subcellular fractions with regard their superoxide dismutase (SOD) activities. The determination of SOD activities based on the well-known spectrophotometric introduced by McCord & Fridovich [(1969) J. Biol. Chem. 244, 6049-6055], partially succinylated (3-carboxypropionylated) rather than native ferricytochrome c as indicating scavenger. Partial succinylation cytochrome resulted in minimization interference associated interaction mitochondrial oxidase or reductases. further increase specificity, exclusion interference, gained a consequence high pH 10 enabled analysis samples rich activity fraction presence absence membrane-disrupting detergents. Linear relationships for dependence protein concentration were obtained rat liver homogenate, microsomal fractions, negligible interference. Furthermore, choosing medium, 4-fold sensitivity compared classical assay, carried out at 7.8, well more precise resolution Cu/Zn-SOD Mn-SOD 2 mM-KCN ratio Cu/Zn-SOD, such mitochondria. complete trapping O2.- radicals, which feasible application simple equation derived calculation appropriately defined units from single experiment.
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H Kuthan, V Ullrich, R W Estabrook, A quantitative test for superoxide radicals produced in biological systems. Biochemical Journal. ,vol. 203, pp. 551- 558 ,(1982) , 10.1042/BJ2030551
D D Tyler, Polarographic assay and intracellular distribution of superoxide dismutase in rat liver. Biochemical Journal. ,vol. 147, pp. 493- 504 ,(1975) , 10.1042/BJ1470493
L. Flohé, F. ötting, Superoxide dismutase assays. Methods in Enzymology. ,vol. 105, pp. 93- 104 ,(1984) , 10.1016/S0076-6879(84)05013-8
Donald G. Lindmark, Miklós Müller, Superoxide Dismutase in the Anaerobic Flagellates, Tritrichomonas foetus and Monocercomonas sp Journal of Biological Chemistry. ,vol. 249, pp. 4634- 4637 ,(1974) , 10.1016/S0021-9258(19)42465-4
Richard A. Weisiger, Irwin Fridovich, Superoxide Dismutase: ORGANELLE SPECIFICITY Journal of Biological Chemistry. ,vol. 248, pp. 3582- 3592 ,(1973) , 10.1016/S0021-9258(19)43969-0
Joe M. McCord, Irwin Fridovich, Superoxide Dismutase AN ENZYMIC FUNCTION FOR ERYTHROCUPREIN (HEMOCUPREIN) Journal of Biological Chemistry. ,vol. 244, pp. 6049- 6055 ,(1969) , 10.1016/S0021-9258(18)63504-5
Angelo Azzi, Cesare Montecucco, Christoph Richter, The use of acetylated ferricytochrome c for the detection of superoxide radicals produced in biological membranes. Biochemical and Biophysical Research Communications. ,vol. 65, pp. 597- 603 ,(1975) , 10.1016/S0006-291X(75)80188-4
Marvin L. Salin, Eugene D. Day, James D. Crapo, Isolation and characterization of a manganese-containing superoxide dismutase from rat liver Archives of Biochemistry and Biophysics. ,vol. 187, pp. 223- 228 ,(1978) , 10.1016/0003-9861(78)90027-9
Bruce L. Geller, Dennis R. Winge, A method for distinguishing Cu,Zn- and Mn-containing superoxide dismutases Analytical Biochemistry. ,vol. 128, pp. 86- 92 ,(1983) , 10.1016/0003-2697(83)90348-2
B.F. Van Gelder, E.C. Slater, The extinction coefficient of cytochrome c Biochimica et Biophysica Acta. ,vol. 58, pp. 593- 595 ,(1962) , 10.1016/0006-3002(62)90073-2