The activity and kinetics of pyruvate kinase in hypoxic newborns.
Pyruvate kinase (PK) plays a key role in erythrocytes, which obtain most of their energy from glycolysis. This study investigated erythrocyte energy metabolism in hypoxic newborns, measuring pyruvate kinase activity, kinetic, and ATP levels in hypoxia. Forty-nine babies who had cord pH value lower than 7.2 and Apgar scores lower than 7 in the first minute were accepted as the hypoxic group, and 48 babies who had cord pH value higher than 7.2 and an Apgar score higher than 7 in the first minute were taken as controls. The erythrocyte mean PK activity was found to be lower (16.9+/-8.5 [5.8-47.9] EU/gHb) in the hypoxic group than the control group (21.3+/-10.9 [3.9-44.3] EU/gHb) (p<.05). The mean ATP value of hypoxic group was higher (19.2+/-11.3 [3.9-37.6] mM) compared to control group (13.8+/-7.16 [3.9-28.7] mM). In the kinetic study, with different ADP concentrations in the control group, the substrate amount (Km) that is needed to reach the half-maximum of enzyme activity (Vmax=27.7 Eu/gHb) was found to be 2.70 mM, but it was 1.47 mM to reach Vmax (22.7 Eu/gHb) in the hypoxic group. Vmax was 41.67 Eu/gHb and Km was 8.33 mM in the control group at different PEP concentrations, whereas Vmax was 21.7 Eu/gHb and Km was 0.89 mM in the hypoxic group. Increase in the ATP level while ePK activity decreases, suggesting that glycolysis increases in hypoxia. In the kinetic study, the substrate amount needed for reaching the half-maximum of enzyme activity was less in the hypoxic group, probably suggests that pyruvate kinase increases glycolysis by increasing its affinity to the substrates. In this way, erythrocytes may gain the energy required for oxygen delivery to tissues and maintaining ion gradient. This arrangement possibly proceeds from sygmoidal structure of pyruvate kinase.
Narli
Nejat
N
Department of Pediatrics, Division of Neonatology, Cukurova University, Faculty of Medicine, Adana, Turkey. nnarli@cu.edu.tr
Satar
Mehmet
M
Kayrin
Levent
L
Yapicioğlu
Hacer
H
Ozlü
Ferda
F
Budgayci
Resul
R
D000244:Adenosine Diphosphate / Q000032:analysis; D000255:Adenosine Triphosphate / Q000032:analysis; D004912:Erythrocytes / Q000201:enzymology*; D005260:Female; D006019:Glycolysis; D006801:Humans; D000860:Hypoxia / Q000097:blood / Q000201:enzymology*; D007231:Infant, Newborn; D007700:Kinetics; D008297:Male; D010100:Oxygen / Q000378:metabolism; D011770:Pyruvate Kinase / Q000032:analysis*
D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D011770:Pyruvate Kinase; D010100:Oxygen