Download Biomembranes - Part D: Biological Oxidations by Sidney Fleischer, Lester Packer PDF

By Sidney Fleischer, Lester Packer

The significantly acclaimed laboratory normal, Methods in Enzymology, is without doubt one of the so much hugely revered courses within the box of biochemistry. due to the fact that 1955, each one quantity has been eagerly awaited, usually consulted, and praised via researchers and reviewers alike. The sequence includes a lot fabric nonetheless appropriate at the present time - actually a vital e-book for researchers in all fields of lifestyles sciences

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2ill, 2105 (1976). 3 b " From W. G. Hanstein, K. A. Davis, M. A. G h a l a m b o r , and Y. Hatefi, 10, 2517 (1971). 65 mM PMS. (mg protein) -1 at 38 °. a At 3 mM ferricyanide. e Moles of s u c c i n a t e oxidized min -1 (mole e n z y m e ) -1 at 38 °. Biochemistry (PMS) and 2,6-dichloroindophenol (DCIP) is 10-11 × 103. The same value is obtained for the turnover number of succinate dehydrogenase in complex II and in submitochondrial particles treated with 2-thenoyltrifluoroacetone (TTFA) to inhibit ubiquinone reduction.

Complex II reacts with ubiquinone, ferricyanide, and phen- 22 ELECTRON TRANSFER COMPLEXES [5] azine methosulfate as electron acceptors, and recombines with the appropriate complexes of the electron transport system to reconstitute highly active succinate-cytochrome c reductase and succinoxidase particles. Purification Procedure The preparation is carried out essentially according to published procedures, 1"2 but with modifications that result in better reproducibility. 3 The pH values of buffers, deoxycholate, and cholate solutions are adjusted at 200-25 °.

Haavik, L. R. Fowler, and D. E. Griffiths, J. Biol. Chem. 237, 2661 (1962). 4 C. R. Hartzell and H. Beinert, Biochim. Biophys. Acta 368, 318 (1974). 5 A. G. Gornall, C. J. Bardawill, and M. M. David, J. Biol. Chem. 177, 751 (1949). 42 ELECTRONTRANSFERCOMPLEXES [8] Source Material. Beef heart mitochondria, prepared by the method of Crane et al. 25 M sucrose and stored overnight at - 2 0 °. Step 1. Washing of Mitochondria. The frozen mitochondrial suspension is thawed in a room-temperature water bath and homogenized with a glass-Teflon homogenizer.

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