The Effect of Long-Term In Vitro Incubation on the Intermediates of Red Cell Metabolism |
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| Authors: | R. J. Henderson Jr G. C. Mills |
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| Affiliation: | Department of Biochemistry and Molecular Biology, Louisiana State University School of Medicine, Shreveport, Louisiana, and the Division of Biochemistry, Department of Human Biological Chemistry and Genetics, The University of Texas Medical Branch, Galveston, Texas |
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| Abstract: | Whole human blood was incubated for periods of up to 70 hours at 37 C under conditions that would cause minimal modifications in normal metabolic controls. Insofar as possible, the conditions utilized were similar to those found in vivo . Control of pH within the limits of 7.2 to 7.5 was achieved by the addition of isotonic Tris buffer or isotonic TES buffer. Glucose, inorganic phosphate, adenine, and pyruvate were added as needed throughout the incubation period. The ADP/ATP ratio and levels of ATP, total adenine nucleotides, 2,3-diphosphoglycerate and other metabolic intermediates were maintained near those of fresh erythrocytes. The maintenance and control of red blood cell adenine nucleotide levels are discussed, with particular emphasis on factors affecting the rate of incorporation of adenine into adenine nucleotides. |
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