Terminal marking of avian triosephosphate isomerases by deamidation and oxidation.
Triosephosphate isomerase (TPI) provides an excellent model for terminal marking and protein degradation. Mammalian TPI is terminally modified by deamidation at Asn71-Gly, resulting in unfolding, dissociation, and proteolysis. In contrast, chicken TPI, which contains a lysine at position 71, is terminally modified by the oxidation of Cys126. Thus, both deamidation and oxidation initiate degradation of TPI from different species. To explore the terminal marking in other avians, we have purified the turkey TPI to homogeneity and determined its characteristics. Although the molecular properties of the turkey and chicken TPI were very similar, their tolerances to temperature, oxidants, and alkaline pH were very different. For example, chicken TPI was inactivated 80% in either 10 mM oxidized glutathione or H2O2, whereas 120 mM GSSG had no effect on turkey TPI, and > 120 mM H2O2 was needed for 80% inactivation. Under alkaline conditions that cause rapid deamidation of the mammalian TPI, neither avian enzyme deamidated. Chicken TPI, however, aggregated. Aggregation was reversed by 2-mercaptoethanol. Under prolonged exposure to milder conditions the turkey enzyme was completely inactivated and deamidated at Asn15-Gly. Thus, there are marked differences in the susceptibility of these two avian enzymes to oxidation and deamidation, and their terminal marking mechanisms appear to be different.
Zhang
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Y
Department of Biochemistry and Molecular Biology, University of North Texas Health Science Center, Fort Worth 76107.
Yüksel
K U
KU
Gracy
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RW
D000595:Amino Acid Sequence; D000818:Animals; D002645:Chickens; D008623:Mercaptoethanol / Q000494:pharmacology; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D011489:Protein Denaturation; D016415:Sequence Alignment; D014305:Triose-Phosphate Isomerase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D014421:Turkey
D008623:Mercaptoethanol; D014305:Triose-Phosphate Isomerase