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51.
Myoglobin: cytochrome b5 interactions and the kinetic mechanism of metmyoglobin reductase 总被引:2,自引:0,他引:2
D J Livingston S J McLachlan G N La Mar W D Brown 《The Journal of biological chemistry》1985,260(29):15699-15707
Metmyoglobin (metMb) reduction by metMb reductase from heart muscle requires cytochrome b5 as electron-transfer mediator. The existence of a metMb-ferrous cytochrome b5 complex is demonstrated by mutual perturbation of the proteins' respective electrophoretic titration curves between pH 4 and 7. The same technique shows a preferential binding of cytochrome b5 over metMb by the enzyme. The paramagnetic hyperfine shifts in the cytochrome b5 1H NMR spectrum are perturbed by metMb, indicating the formation of a specific bimolecular complex with a 1:1 stoichiometry and a binding constant estimated to be less than 10 microM. The resonances assigned to the cytochrome b5 heme 6-propionate methylene group exhibit the largest complexation shifts. Computer modeling implicates lysines 47, 50, and 98 of metMb as contact points with cytochrome b5 carboxylate residues 43, 44, 60, and heme 6-propionate. The mechanism of the enzymatic reduction establishes metMb reductase as an NADH-cytochrome b5 oxidoreductase. Cytochrome b5 is reduced at near diffusion-controlled rates by the enzyme with a turnover number of 1000 min-1 X Km for the cytochrome is 0.9 microM versus 100 microM reported for the erythrocyte enzyme. Ferrous cytochrome b5 then reduces metMb nonenzymatically with an apparent rate constant of 4.9 X 10(4) M-1 min-1 X Acetylation of metMb, which does not affect its oxygen affinity or chemical reduction, renders it a poor substrate for enzymatic reduction. This study suggests a function for the three exterior lysine residues conserved in all mammalian myoglobin sequences: they are contact points for complexation with cytochrome b5. 相似文献
52.
Among the apoA-I-containing lipoproteins isolated by selected-affinity immunosorption from human serum and plasma, we have identified a subpopulation which, unlike the bulk of high density lipoproteins, has pre-beta electrophoretic mobility. This pre-beta subpopulation can be observed directly in fresh plasma by immunoelectrophoresis. It contains phospholipid and free and esterified cholesterol, but protein constitutes 90% of its mass. Apolipoprotein A-I is the predominant apolipoprotein in this subpopulation; apolipoprotein A-II and the B lipoproteins are not detected. The protein moiety of this subpopulation exhibits markedly lower helicity than that of high density lipoproteins isolated by ultracentrifugation. 相似文献
53.
G Gravante F Sorbera M Sabatino V La Grutta 《Bollettino della Società italiana di biologia sperimentale》1985,61(2):249-254
In chronic free-moving cats unilateral, low frequency, long duration stimulation of the caudate nucleus causes a significant reduction of food intake. The effects are greatest at the beginning of the stimulation. On cessation of stimulation the animals take enough food to compensate for that lost during the time of stimulation. Results are discussed either considering interrelationships between striatum and lateral hypothalamus, either regarding recent viewpoints distinguishing two striatal compartments: dorsal or "non limbic" striatal region and ventral or "limbic" striatal region. 相似文献
54.
The protein synthesis activity of heart, skeletal muscle and liver polysomes from isoprotenerol-treated and control hamsters has been compared in an in vitro non-inititating translation system. Heart and skeletal muscle polysomes from treated hamsters were less active than controls and required a higher magnesium concentration for optimal protein synthesis. These results suggest that there is a conformational modification in heart and skeletal muscle ribosomes from isoprotenerol-treated hamsters. No such change was observed with ribosomes from the liver of isoproterenol-treated hamsters. 相似文献
55.
56.
Summary Expression of the anchorage-independent transformed phenotype in BHK 21/13 cells generally behaves as a recessive trait. When
chemically induced and spontaneously arising transformants are fused to the nontransformed parent line, transformation is
initially suppressed, reappearing after extended growth of the hybrids. In this paper, complementation for the expression
of anchorage independence was sought among a large group of such transformants, all independently derived from BHK 21/13 cells.
Tumorigenicity studies on selected hybrids and parental lines indicated that the in vitro trait of anchorage independence
is an accurate indicator of in vivo neoplasia for these cells. Seventeen of the 18 clones tested did not complement one or
more of three tester strains. This result indicates that anchorage independence arose in these clones as a result of lesions
in the same genetic function and suggests that the final step in the progressive changes of carcinogenesis may frequently
be restricted to lesions at a single locus.
This investigation was supported by National Institutes of Health grant CA27306. 相似文献
57.
58.
Jef D. Boeke Francois La Croute Gerald R. Fink 《Molecular & general genetics : MGG》1984,197(2):345-346
Summary Mutations at the URA3 locus of Saccharomyces cerevisiae can be obtained by a positive selection. Wild-type strains of yeast (or ura3 mutant strains containing a plasmid-borne URA3
+ gene) are unable to grow on medium containing the pyrimidine analog 5-fluoro-orotic acid, whereas ura3
– mutants grow normally. This selection, based on the loss of orotidine-5-phosphate decarboxylase activity seems applicable to a variety of eucaryotic and procaryotic cells. 相似文献
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