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Glycolysis of Small Amounts of Glucose by Ehrlich Ascites Carcinoma Cells¹

Department of Biological Chemistry, School of Medicine, University of California, Los Angeles, California, and Department of Biochemistry, Northwestern University Medical School, Chicago, Illinois

Suspensions of about 3 volumes % of Ehrlich ascites carcinoma cells in 54 mM phosphate buffer, pH 7.4, were aerobically incubated. Changes were measured in glucose, ATP,⁴ and glycolytic products during the 1st 3 min after addition of as little as 27 µM glucose (0.9 µmoles/ml of cells).

During the 1st 60 sec, lactic acid, fructose diphosphate, and dihydroxyacetone phosphate almost completely account for the glucose utilized. Addition of 3.5 µmoles of glucose/ml of cells at 37°C elicits the following sequence: Maximum fructose diphosphate accumulation, 0.3–0.4 min; maximum dihydroxyacetone phosphate accumulation, 0.5–0.6 min; the maximum rate of theoretical glycolytic ATP synthesis, 0.4–0.6 min; and the end of glycolysis, 1.5–2.0 min. The maximum rate of glycolytic ATP synthesis, estimated from product accumulation, ranged from 4 to 8 µmoles of ATP/ml of cells/min.

A variation of only 4–10 µmoles of lactate/ml of cells/min was observed in the initial rate of lactate accumulation, although the amount of glucose added ranged from 0.9 to 42 µmoles/ml of cells. The average initial rate was about 70% higher than the steady-state rate of lactate accumulation observed 5 min or more after addition of high concentrations of glucose.

The amount of glucose accounted for in terms of the total lactate accumulated increases below 4 µmoles of glucose/ml of cells. The maximum fructose diphosphate accumulated increases as a function of the glucose added.

¹ The initial phases of this work were supported by grants-in-aid from the National Cancer Institute, USPHS (C-2006) and the Cancer Coordinating Committee, University of California; the extensions of the work were supported by a grant from the NIH (GM-11580-01).

⁴ Abbreviations used: ADP, adenosine-5'-diphosphate; ATP, adenosine-5'-triphosphate; DHAP, dihydroxyacetone phosphate; DPN, diphosphopyridine nucleotide; DPNH, reduced diphosphopyridine nucleotide; DHAP, dihydroxyacetone phosphate; FDP, fructose-1,6-diphosphate; G, glucose; L, lactic acid; PCA, perchloric acid; PGA, monophosphoglyceric acid; TPN, triphosphopyridine nucleotide; Tris, tris(hydroxymethyl)aminomethane.

² Present address: Department of Biochemistry, Northwestern University Medical School, Chicago, Illinois.

³ Present address: California College of Medicine, Los Angeles, California.

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