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Correlation of Glycolytic and Respiratory Events after Addition of a Small Amount of Glucose to Ehrlich Ascites Carcinoma¹

Department of Biochemistry, Northwestern University Medical School, Chicago, Illinois

The accumulations of lactate, pyruvate, and several phosphorylated compounds were measured in a suspension containing 1.8 volumes % of Ehrlich ascites tumor cells in 54 mM phosphate (pH 7.35) at 30°C after addition of 0.08 mM glucose. The ATP² level increased from 0.3 to 2.1 µmoles/ml of cells during a period of 120 sec. All measured phosphorylated glycolytic intermediates rose to maxima and then declined to the endogenous level by 120–150 sec except phosphopyruvate, which exhibited a distinct decline during the 1st 30 sec and then rose again.

The rate of oxygen consumption, as estimated with a bare, vibrating-reed, platinum-electrode polarograph, decreased during the 1st 45 sec, remained low until 150 sec, and then increased to a plateau that was reached by 270 sec. Additional experiments confirmed that recovery of respiration from inhibition lagged behind the cessation of glycolysis.

The sum of glycolytic and respiratory ATP synthesis (estimated by assuming complete coupling) gave a curve that rose during the 1st 30 sec, fell sharply between 90 and 150 sec, and rose again between 150 and 270 sec. The results are interpreted to mean that glycolytically synthesized ATP is more immediately available for cellular maintenance than oxidatively generated ATP.

¹ This work was supported by a grant (GM-11580-01) from the NIH.

² Abbreviations used: ADP, adenosine-5'-diphosphate; ATP, adenosine-5'-triphosphate; DHAP, dihydroxyacetone phosphate; DPN, diphosphopyridine nucleotide; DPNH, reduced diphosphopyridine nucleotide; FDP, fructose-1,6-diphosphate; G, glucose; L, LACT, lactic acid; PCA, perchloric acid; PEP, phosphoenol pyruvic acid; PGA, monophosphoglyceric acid; TP, total triose phosphate.

Received 3/22/65. Revised 8/26/65.