title: Conversion of amino acid residues in proteins and amino acid homopolymers to carbonyl derivatives by metal-catalyzed oxidation reactions.
abstract: A number of metal-catalyzed oxidation (MCO) systems mediate the oxidative inactivation of enzymes. This oxidation is accompanied by conversion of the side chains of some amino acid residues to carbonyl derivatives (for review, see Stadtman, E. R. (1986) Trends Biochem. Sci. 11, 11-12). To identify the amino acid residues which are sensitive to MCO oxidation, several enzymes/proteins and amino acid homopolymers were exposed to various MCO systems. The carbonyl groups which were formed were converted to their corresponding 3H-labeled hydroxy derivatives. After acid hydrolysis, the labeled free amino acids were separated by ion exchange chromatography. Each protein or polymer gave rise to several different labeled amino acids. The elution profiles of the labeled amino acids obtained from preparations of Escherichia coli glutamine synthetase which had been oxidized by MCO systems comprised of either Fe(II)/O2 or ascorbate/Fe(II)/O2 both in the presence and absence of EDTA were qualitatively the same. From a comparison of the elution profiles of labeled amino acids from various proteins with those obtained from homopolymers, it is evident that the side chains of histidine, arginine, lysine, and proline are particularly sensitive to oxidation by the MCO systems. This conclusion is supported also by direct amino acid analysis of acid hydrolysates which shows that the oxidation of glutamine synthetase, enolase, and phosphoglycerate kinase is associated with the loss of at least 1 histidine residue per subunit. From the results of studies with homopolymers, it is apparent that glutamic semialdehyde is a major product of both proline and arginine residues. In addition, hydroxyproline and unlabeled glutamic acid were identified among the hydrolysis products of oxidized poly-L-proline, and unlabeled aspartic acid was identified as a product of poly-L-histidine oxidation.
authors.affiliation: Laboratory of Biochemistry, National Heart, Lung, and Blood Institute, Bethesda, Maryland 20892.
authors.forename: A
authors.initials: A
authors.lastname: Amici
authors.forename: R L
authors.initials: RL
authors.lastname: Levine
authors.forename: L
authors.initials: L
authors.lastname: Tsai
authors.forename: E R
authors.initials: ER
authors.lastname: Stadtman
mesh_terms: D000596:Amino Acids / Q000378:metabolism*; D001120:Arginine / Q000378:metabolism; D001894:Borohydrides; D004492:Edetic Acid / Q000378:metabolism / Q000494:pharmacology; D005290:Ferric Compounds / Q000378:metabolism; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism; D005974:Glutamate-Ammonia Ligase / Q000378:metabolism; D006639:Histidine / Q000378:metabolism; D006859:Hydrogen / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism; D008239:Lysine / Q000378:metabolism; D008345:Manganese / Q000494:pharmacology; D008670:Metals / Q000378:metabolism*; D006899:Mixed Function Oxygenases / Q000378:metabolism; D010084:Oxidation-Reduction; D010100:Oxygen / Q000378:metabolism; D011108:Polymers; D011392:Proline / Q000378:metabolism; D011506:Proteins / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D001894:Borohydrides; D005290:Ferric Compounds; D008670:Metals; D011108:Polymers; D011506:Proteins; C024823:ferric sulfate; D008345:Manganese; D006639:Histidine; D006859:Hydrogen; C025364:sodium borohydride; D001120:Arginine; D011392:Proline; D004492:Edetic Acid; D006861:Hydrogen Peroxide; D006899:Mixed Function Oxygenases; D005954:Glucosephosphate Dehydrogenase; D005974:Glutamate-Ammonia Ligase; D008239:Lysine; D010100:Oxygen