Soluble cytochrome b5 reductase from human erythrocytes

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Soluble cytochrome b 5 reductase from human erythrocytes.

I. An enzyme that catalyzes the reduction of erythrocyte cytochrome b 5 has been isolated from the supernatant fraction of erythrocyte hemolysates by chromatography on DEAE-cellulose, Amberlite CG-5 o, and Bio-Gel P-6o. 2. Erythrocyte cytochrome b 5 reductase has been shown to contain FAD. Incubation of the reductase in the absence of EDTA results in both the appearance of flavin fluorescence a...

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Membrane-bound cytochrome b5 reductase (methemoglobin reductase) in human erythrocytes. Study in normal and methemoglobinemic subjects.

In this study we present evidence that in human erythrocytes NADH-cytochrome b5 reductase (methemoglobin reductase) is not only soluble but also tightly bound to the membrane. The membrane methemoglobin reductase-like activity is unmasked by Triton X-100 treatment, and represents about half of the total activity in the erythrocytes. Like the amphiphilic microsomal-bound cytochrome b5 reductase,...

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Properties of NADH-cytochrome-b5 reductase from human neutrophils.

An NADH-ferricyanide reductase activity of ca. 170 nmole ferricyanide reduced/min/10(7) cells is present in the membrane fraction of human neutrophils. This membrane-bound activity constitutes ca. 85% of the total NADH-ferricyanide reductase activity that is present in these cells. The enzyme(s) readily utilize(s) purified cytochrome-b5 from beef liver as an electron acceptor. No other physiolo...

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Characterization of the purified NADH-cytochrome b5 reductase of human erythrocytes as a FAD-containing enzyme.

NADH-cytochrome b5 reductase of normal human erythrocytes was purified by procedures including affinity chromatography on Blue-Sepharose to an electrophoretically homogeneous protein. The purified enzyme was judged to be a typical flavoprotein based on its absorption spectrum (absorption maxima, 272, 390, and 462 nm; shoulders, 373 and 488 nm) and flavin content (1 mol of FAD/mol of enzyme). Th...

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Improved determination of cytochrome b5 in human erythrocytes.

A sensitive, precise enzymic/spectrophotometric method for determining cytochrome b5 in small amounts of blood is described. Mean values for healthy individuals, ages 20 to 70 years, were 0.26 (SD 0.03) mumol per liter of erythrocytes or 0.87 (SD 0.14) nmol per gram of hemoglobin. We believe the assay is preferable to methods described hitherto, primarily because of its high sensitivity.

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ژورنال

عنوان ژورنال: Biochimica et Biophysica Acta (BBA) - Bioenergetics

سال: 1972

ISSN: 0005-2728

DOI: 10.1016/0005-2728(72)90024-2