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The youngest and oldest erythrocytes of normal human blood were segregated by buoyant density gradients of BSA. PK in hemolysates of older cells was observed to differ significantly from that in the younger cells by virtue of lowered optimal activity, increased pH optimum, decreased electrophoretic mobility, and elevated Michaelis constant for PEP. The latter was correctable to normal values by small amounts of the activator, fructose diphosphate. The data suggest that molecular changes in this enzyme associated with in vivo cell aging may be one pertinent mechanism of senescence in normal human erythrocytes.
Abbreviations:PK (pyruvate kinase), Km (apparent Michaelis-Menten constant), PEP (phosphoenolpyruvate), Vmax (maximum velocity of enzymatic reaction under optimal conditions), FDP (fructose-1,6-diphosphate), BSA (bovine serum albumin), RCF (relative centrifugal force), G-6-PD (glucose-6-phosphate dehydrogenase), ADP (adenosine diphosphate), TEA (triethanolamine hydrochloride), U (enzyme units), NADH (reduced nicotine adenine dinucleotide (formerly DPNH)), NAD (oxidized nicotine adenine dinucleotide (formerly DPN)), LDH (lactate dehydrogenase)
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Accepted: May 22, 1970
Received: March 19, 1970With the technical assistance of KLAUS K. KÜRSCHNER and MARY LU WILSON
☆Supported by grants from the Veterans Administration and the United States Public Health Service.
☆☆Miss Ruth Exley kindly provided assistance with manuscript preparation.
© 1970 Published by Elsevier Inc.