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Effect of temperature on red cell ATP
117
Clinica Chimica Acfu, 199 (1991) 717, 118 8 1991 Elsevier Science Publishers B.V. 0009-8981/91/$03.50
CCA 04985
Letter to the Editor
Effect of temperature on red cell ATP Dear Editor, Simmons et al. report that ATP activities decline rapidly in hepa~nized blood stored at 4”C, but more slowly at room temperature [l]. We first observed this phenomenon over 25 years ago [2] and were able to show that it is a consequence of the rise that occurs in the pH of heparinized blood when it is cooled to 4’ C [3,4], a phenomenon first described by Rosenthal in 1948 [5]. In investigating this phenomenon we found that the loss of ATP could be prevented by o~tt~ng glucose from the suspending medium 141. The activity of phosphofructokinase and hexokinase is greatly increased by the alkaline pH and thus ATP is more rapidly dephosphorylated to ADP. The high pH also serves to displace the lactate dehydrogenase equilibrium in the direction of pyruvate and NADH, so that a functional block occurs at the glyceraldehyde phosphate dehydrogenase step. Thus, substrate is not available for the rephospho~lation of ADP to ATP in the phosphogly~erate kinase and pyruvate kinase steps. The effect of storing heparinized blood in ice for even 10 minutes has an even greater effect on some of the trace intermediates. Fructose diphosphate levels of heparinized blood at 0°C are increased more than three-fold in even 10 minutes 161. Thus, Simmons et al. are quite correct in pointing out that marked metabolic changes occur when blood is cooled. When one takes into account what is known about the effect of pH on red cell metabolism the findings are readily explained. Ernest Beutler Scripps Clinic and Research Foundation La Jolla. CA, USA
References 1 Simmonds HA, Micheli V, Davies PM, McBride MB. Erythrocyte nucleotide stability and plasma hypoxanthine concentrations: Improved ATP stability with short-term storage at room temperature. Clin Chim Acta 1990;192:121-132. 2 Beutler E, Mathai CK. A comparison of normal red cell ATP levels as measured by the firefly system and the hexokinase system. Blood 1967;30:311-320.
Correspondence to: Dr. E. Beutler, Road, La Jolla, CA 92037, USA.
Scripps
Clinic
and Research
Foundation,
10666 N. Torrey
Pines
118 3 Beutler E, Duron 0. Factors influencing the preservation of red cell ATP on storage. Folia Haematol (Leipz) 1965;83:403-409. 4 Beutler E, Duron 0. Effect of pH on preservation of red cell ATP. Transfusion 1965;5:17-24. 5 Rosenthal TB. The effect of temperature on the pH of blood and plasma in vitro. J Biol Chem 1948;173:25-30. 6 Niessner H, Beutler E. Fluorometric analysis of glycolytic intermediates in human red blood cells. Biochem Med 1973:8:123-134.
Clinica Chimica Acfu, 199 (1991) 717, 118 8 1991 Elsevier Science Publishers B.V. 0009-8981/91/$03.50
CCA 04985
Letter to the Editor
Effect of temperature on red cell ATP Dear Editor, Simmons et al. report that ATP activities decline rapidly in hepa~nized blood stored at 4”C, but more slowly at room temperature [l]. We first observed this phenomenon over 25 years ago [2] and were able to show that it is a consequence of the rise that occurs in the pH of heparinized blood when it is cooled to 4’ C [3,4], a phenomenon first described by Rosenthal in 1948 [5]. In investigating this phenomenon we found that the loss of ATP could be prevented by o~tt~ng glucose from the suspending medium 141. The activity of phosphofructokinase and hexokinase is greatly increased by the alkaline pH and thus ATP is more rapidly dephosphorylated to ADP. The high pH also serves to displace the lactate dehydrogenase equilibrium in the direction of pyruvate and NADH, so that a functional block occurs at the glyceraldehyde phosphate dehydrogenase step. Thus, substrate is not available for the rephospho~lation of ADP to ATP in the phosphogly~erate kinase and pyruvate kinase steps. The effect of storing heparinized blood in ice for even 10 minutes has an even greater effect on some of the trace intermediates. Fructose diphosphate levels of heparinized blood at 0°C are increased more than three-fold in even 10 minutes 161. Thus, Simmons et al. are quite correct in pointing out that marked metabolic changes occur when blood is cooled. When one takes into account what is known about the effect of pH on red cell metabolism the findings are readily explained. Ernest Beutler Scripps Clinic and Research Foundation La Jolla. CA, USA
References 1 Simmonds HA, Micheli V, Davies PM, McBride MB. Erythrocyte nucleotide stability and plasma hypoxanthine concentrations: Improved ATP stability with short-term storage at room temperature. Clin Chim Acta 1990;192:121-132. 2 Beutler E, Mathai CK. A comparison of normal red cell ATP levels as measured by the firefly system and the hexokinase system. Blood 1967;30:311-320.
Correspondence to: Dr. E. Beutler, Road, La Jolla, CA 92037, USA.
Scripps
Clinic
and Research
Foundation,
10666 N. Torrey
Pines
118 3 Beutler E, Duron 0. Factors influencing the preservation of red cell ATP on storage. Folia Haematol (Leipz) 1965;83:403-409. 4 Beutler E, Duron 0. Effect of pH on preservation of red cell ATP. Transfusion 1965;5:17-24. 5 Rosenthal TB. The effect of temperature on the pH of blood and plasma in vitro. J Biol Chem 1948;173:25-30. 6 Niessner H, Beutler E. Fluorometric analysis of glycolytic intermediates in human red blood cells. Biochem Med 1973:8:123-134.