title: Affinity labels for the anion-binding site in ovotransferrin.
abstract: Bromopyruvate, a known alkylating agent, has previously been reported to function as an affinity label for the anion-binding site in the iron-binding protein ovotransferrin [Patch, M.G., & Carrano, C. J. (1982) Biochim. Biophys. Acta 700, 217-220]. However, the present results indicate that hydroxypyruvate also functions in an almost identical manner, which implies that alkylation of a susceptible nucleophile cannot be the mechanism responsible for the covalent attachment of the anion. Model complexes and amino acid analysis of labeled ovotransferrin suggest that initial Schiff base formation, followed by reduction of the imine bond between the affinity anion and a lysine within the locus of the anion-binding site, accounts for the irreversible labeling. As expected, the covalently attached anions render the iron in the ovotransferrin-iron-anion ternary complex much more resistant to loss at low pH. It is proposed that the covalently labeled protein be used to test the hypothesis that iron removal from transferrin occurs by protonation and loss of the anion in low-pH lysosomal vesicles.
authors.affiliation: Department of Chemistry, University of Vermont, Burlington 05405.
authors.forename: C T
authors.initials: CT
authors.lastname: Bailey
authors.forename: M G
authors.initials: MG
authors.lastname: Patch
authors.forename: C J
authors.initials: CJ
authors.lastname: Carrano
mesh_terms: D000345:Affinity Labels / Q000378:metabolism*; D000838:Anions; D001665:Binding Sites; D003207:Conalbumin / Q000378:metabolism*; D004527:Egg Proteins / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007501:Iron / Q000378:metabolism; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D011485:Protein Binding; D011773:Pyruvates / Q000378:metabolism*; D013053:Spectrophotometry; D014168:Transferrin / Q000378:metabolism
chemical_list: D000345:Affinity Labels; D000838:Anions; D004527:Egg Proteins; D011773:Pyruvates; D014168:Transferrin; D003207:Conalbumin; C017092:bromopyruvate; D007501:Iron