Exposure of rat muscle phosphoglycerate kinase to a nonenzymatic MFO system generates the old form of the enzyme.
The occurrence of age-related modifications in enzymes is a well-established symptom of aging that has been explained by several possible mechanisms including the oxidation of amino acid residues by mixed-function oxidation (MFO) systems. In the present study native old phosphoglycerate kinase was compared with young enzyme which had been modified by oxidation with ascorbate: FeCl3 followed by reduction. The comparison was done by monitoring the rates of heat denaturation of these enzyme forms, as well as their inactivation by trypsin. A remarkable similarity between the old and treated young enzyme was revealed, while native young phosphoglycerate kinase was inactivated with a different rate. Extensive unfolding followed by refolding converted both old and MFO-treated young phosphoglycerate kinase to species which greatly resemble the native young enzyme in their heat inactivation kinetics. These results demonstrate that the exposure of phosphoglycerate kinase to the mixed-function oxidation system introduces some modifications which are not reversed by subsequent enzyme reduction and which resemble those found in the native old enzyme. This mechanism, therefore, may account for the aging of phosphoglycerate kinase in vivo.
Zhou
J Q
JQ
Institute of Gerontology, University of Michigan, Ann Arbor.
Gafni
A
A
D000375:Aging / Q000378:metabolism*; D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D002712:Chlorides; D003545:Cysteine / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D005260:Female; D005290:Ferric Compounds / Q000494:pharmacology; D006358:Hot Temperature; D008623:Mercaptoethanol / Q000494:pharmacology; D009132:Muscles / Q000201:enzymology*; D010084:Oxidation-Reduction; D010735:Phosphoglycerate Kinase / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D011487:Protein Conformation; D051381:Rats; D011919:Rats, Inbred Strains; D014357:Trypsin / Q000494:pharmacology
D002712:Chlorides; D005290:Ferric Compounds; D008623:Mercaptoethanol; D010735:Phosphoglycerate Kinase; D014357:Trypsin; D003545:Cysteine; D001205:Ascorbic Acid; C024555:ferric chloride