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Sahabeler Ansiklopedisi Pdf Indir Free
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The enzyme NADH dehydrogenase has been isolated from pig liver mitochondria by the aid of a number of purification procedures (isoelectric focusing, DEAE-cellulose, hydroxylapatite, Sephadex G-100 and hydroxyapatite). The purified enzyme is of uniform molecular weight (160,000) on polyacrylamide gels and has a pH-optimum of 6.8 and a Michaelis constant for the oxidation of NADH of 28 microM. The enzyme combines with FAD at a ratio of 1:1, i.e. it contains 1 mole FAD/160,000 daltons. The enzyme has been studied in more detail with respect to substrate specificity and subunit structure. The enzyme was active with both NADH and NAD+ as substrates. For both substrates the Km for cytochrome c is in the micromolar range. FAD has no effect on the affinity for NAD+ but inhibits the oxidation of both NADH and NAD+. After oxidative breakdown of FAD the enzyme loses its FAD and is inactive. The holoform contains heme and the iron in the d orbitals. The EPR spectrum of the complexed holoenzyme shows the typical spectrum of a low spin ferric heme group. In the presence of NAD+, the heme undergoes a conversion into a high spin species which is a characteristic of the low spin Fe2+ heme of cytochrome b5. Case: 12-40887 Document: 00512212087 Page: 1 Date Filed: 04/17/2013
IN THE UNITED STATES COURT OF APPE
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