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We have studied the affinity of methylmalonyl CoA mutase for its required cofactor, adenosylcobalamin, in extracts of control and mutant human cultured fibroblasts. Control enzyme has an apparent Km for adenosylcobalamin of 6–7 × 10?8 M. Five mutant cell lines from patients with methylmalonicacidemia due to a mutase apoenzyme defect were studied. Three have undetectable mutase activity (<0.15% of control) at all cofactor concentrations. Two others, however, have markedly altered Km's for adenosylcobalamin of 2.8 × 10?4 M and 1.7 × 10?5 M. These mutant lines synthesize adenosylcobalamin normally and, by complementation analysis, are genetically identical to all other mutase apoenzyme mutants tested. We conclude that the mutase deficiency in these two cell lines results from structurally altered mutase apoenzymes with markedly reduced affinities for adenosylcobalamin. 相似文献
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G M Greig S Parikh J George V E Powers H F Willard 《Cytogenetics and cell genetics》1991,56(3-4):144-148
A 340-bp EcoRI fragment of alpha satellite DNA from human chromosome 12 has been isolated and used in molecular cytogenetic and genetic studies. The clone, pSP12-1, detects tandemly repeated 1.4-kb repeat units at the centromeric region of chromosome 12. By fluorescence in situ hybridization, biotinylated pSP12-1 is highly specific for chromosome 12 and has been used to confirm an i(12p) in a case of Pallister-Killian syndrome, both in metaphase spreads and in interphase nuclei. A dominant DNA polymorphism for the centromeric D12Z3 locus is detected with the enzyme TaqI. In addition, a high frequency of D12Z3 array length polymorphisms can be detected using pulsed-field gel electrophoresis. The D12Z3 array has been measured by pulsed-field gel electrophoresis to span approximately 2,250-4,300 kb at the centromeric region of chromosome 12. 相似文献
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John C. Maynard Stuart M. Mertz Jr. Charles J. Arntzen Willard W. Payne 《American journal of botany》1974,61(6):580-584
A study of a mutant variety of Zea mays (ON8147) revealed that the mutant plants, in contrast with normal maize plants, do not exhibit a light-induced increase in the rate of transpiration, and that the ontogeny of the stomatal complex is abnormal. In later stages of differentiation, the guard cells of mutant plants deteriorate, leaving the mature stomata with only the two subsidiary cells. The subsidiary cells in stomata of mutant leaves are similar to those of normal leaves with respect to their capacity to accumulate K+ in the dark, but they do not lose K+ in the light, as do subsidiary cells of stomata of nonmutant plants. It is suggested that impairment of guard cell function causes death of the mutant plant seedlings primarily by restricting CO2 entry into the leaf. 相似文献
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The present paper describes a new spectrophotometric assay for HGPRTase activity which is more rapid than and as sensitive as the isotopic assays for this enzyme and which avoids the use of high-voltage electrophoresis and liquid scintillation counting. 相似文献
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