Studies on the mechanism of acetaldehyde-mediated inhibition of aspartate aminotransferase (GOT) in human erythrocytes.
The mechanism of acetaldehyde-mediated GOT inhibition was studied in human red cells. In the GOT assay mix, acetaldehyde competitively inhibits activation of apoGOT by pyridoxal phosphate and pyridoxamine phosphate. However, Ki values are 100-1000 times greater than Km values for these B6 vitamers. Moreover, incubation of undiluted lysates with acetaldehyde at 37 degrees inhibits GOT activity without increasing apoGOT levels and without altering affinity of apoGOT for either B6 coenzyme. In undiluted lysates, inhibition is not prevented by disulfiram. However, incubation at 4 degrees prevents both acetaldehyde metabolism and GOT inhibition while preincubation with NaF prevents GOT inhibition without affecting acetaldehyde disappearance. The effect of NaF is completely reversed by pyruvate but only partially reversed by NADH. Glyceraldehyde 3-phosphate, the only glycolytic intermediate which directly inhibits GOT, does not reverse the NaF effect. Thus, inhibition of GOT by acetaldehyde (a) requires nonoxidative metabolism of acetaldehyde and (b) is not mediated either by glycolytic substrates or by impaired binding of B6 vitamers to the GOT apoenzyme. Since NaF had no effect on a lysate deficient in glucose 6-phosphate dehydrogenase, the hexose monophosphate shunt may play a role in acetaldehyde-mediated GOT inhibition.
Solomon
L R
LR
Department of Medicine, Veterans Administration Medical Center, West Haven, Connecticut 06516.
Crouch
J Y
JY
D000079:Acetaldehyde / Q000494:pharmacology*; D000255:Adenosine Triphosphate / Q000494:pharmacology; D001219:Aspartate Aminotransferases / Q000037:antagonists & inhibitors* / Q000097:blood; D003067:Coenzymes / Q000097:blood; D004912:Erythrocytes / Q000201:enzymology*; D005955:Glucosephosphate Dehydrogenase Deficiency / Q000097:blood / Q000201:enzymology; D006019:Glycolysis; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D011725:Pyridines / Q000494:pharmacology; D012969:Sodium Fluoride / Q000494:pharmacology
D003067:Coenzymes; D011725:Pyridines; D000255:Adenosine Triphosphate; D012969:Sodium Fluoride; D001219:Aspartate Aminotransferases; D000079:Acetaldehyde