title: 1,2-Cyclohexanedione modification of arginine residues in egg-white riboflavin-binding protein.
abstract: 1. Reaction of 1,2-cyclohexanedione with arginine residues of egg white riboflavin-binding protein results in a loss of the binding activity. 2. In borate buffer pH 8.0, with 0.15 M cyclohexanedione, the inactivation proceeds with a pseudo-first-order rate constant 0.084 hr.-1. 3. At least 65% of lost riboflavin binding capacity can be recovered on 12 hr incubation in 0.5 M hydroxylamine pH 7.0. 4. All 5 arginine residues are modified, 2-3 of them seem to react much easier than others. 5. The correlation between modification of arginines and protein inactivation, as analyzed by kinetic and statistical methods, suggests that one of low-reactivity residues is "essential" for riboflavin binding. 6. In the holoprotein, one arginine residue is almost completely protected from 1,2-cyclohexanedione modification. 7. Riboflavin does not dissociate from holoprotein, even on prolongated incubation with the reagent. 8. The protected arginine residue seems to be located in the riboflavin binding pocket of protein macromolecule.
authors.affiliation: Jagiellonian University, Institute of Molecular Biology, Kraków, Poland.
authors.forename: A
authors.initials: A
authors.lastname: Kozik
authors.forename: I
authors.initials: I
authors.lastname: Guevara
authors.forename: Z
authors.initials: Z
authors.lastname: Zak
mesh_terms: D001120:Arginine / Q000032:analysis / Q000502:physiology*; D002352:Carrier Proteins / Q000032:analysis / Q000378:metabolism*; D003510:Cyclohexanes / Q000494:pharmacology*; D003512:Cyclohexanones / Q000494:pharmacology*; D004527:Egg Proteins / Q000378:metabolism*; D026901:Membrane Transport Proteins; D012256:Riboflavin / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D003510:Cyclohexanes; D003512:Cyclohexanones; D004527:Egg Proteins; D026901:Membrane Transport Proteins; C019079:riboflavin-binding protein; C018031:1,2-cyclohexanedione; D001120:Arginine; D012256:Riboflavin