31P NMR measurements of the ADP concentration in yeast cells genetically modified to express creatine kinase.
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.
Brindle
K
K
Department of Biochemistry, University of Oxford, England.
Braddock
P
P
Fulton
S
S
D000244:Adenosine Diphosphate / Q000378:metabolism*; D000818:Animals; D003402:Creatine Kinase / Q000235:genetics* / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D010957:Plasmids; D011817:Rabbits; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic
D000244:Adenosine Diphosphate; D003402:Creatine Kinase