P-31 NMR MEASUREMENTS OF THE ADP CONCENTRATION IN YEAST-CELLS GENETICALLY MODIFIED TO EXPRESS CREATINE-KINASE

被引：23
|
作者：
BRINDLE, K
BRADDOCK, P
FULTON, S
机构：
[1] Department of Biochemistry, University of Oxford, Oxford OX1 3QU, South Parks Road
来源：
BIOCHEMISTRY | 1990年 / 29卷 / 13期
关键词：
D O I：
10.1021/bi00465a021
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
Rabbit muscle creatine kinase has been introduced into the yeast Saccharomyces cerevisiae by transforming cells with a multicopy plasmid containing the coding sequence for the enzyme under the control of the yeast phosphoglycerate kinase promoter. The transformed cells showed creatine kinase activities similar to those found in mammalian heart muscle. 31P NMR measurements of the near-equilibrium concentrations of phosphocreatine and cellular pH together with measurements of the total extractable concentrations of phosphocreatine and creatine allowed calculation of the free ADP/ATP ratio in the cell. The calculated ratio of approximately 2 was considerably higher than the ratio of between 0.06 and 0.1 measured directly in cell extracts. © 1990, American Chemical Society. All rights reserved.
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页码：3295 / 3302
页数：8
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