title: Effects of pH and ionic strength on the binding of egg white riboflavin binding protein with flavins.
abstract: The effects of pH and ionic strength on the equilibrium constants and rate constants (binding and dissociation rate constants) between riboflavin binding protein (RBP) and flavins (riboflavin, 3-carboxymethylriboflavin [CMRF], and FMN) were studied by fluorometry. The equilibrium constant and the binding rate constant between RBP and riboflavin were pH-independent between pH 6 and 9, and both constants were also independent of the ionic strength, while the constants between RBP and CMRF or FMN were dependent on both pH and ionic strength. The dissociation rate constants between RBP and the flavins used here were not so dependent on pH and ionic strength in the pH region 6 to 9, and the patterns of pH profiles as a whole were similar to each other, although the constants for FMN were about 30-60 times larger than those for CMRF or riboflavin. RBP had lower affinity for FMN than for riboflavin in the neutral pH region, which is based on the small binding rate constant and the large dissociation rate constant for FMN. The former is due to an electrostatic repulsion force between negative net charges of RBP and the phosphate group of FMN, and the latter is due to steric interference by the phosphate group of FMN.
authors.affiliation: Department of Obstetrics and Gynecology, Kumamoto University Medical School.
authors.forename: H
authors.initials: H
authors.lastname: Ushijima
authors.forename: H
authors.initials: H
authors.lastname: Okamura
authors.forename: Y
authors.initials: Y
authors.lastname: Nishina
authors.forename: K
authors.initials: K
authors.lastname: Shiga
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000032:analysis*; D002645:Chickens; D004529:Egg White / Q000032:analysis*; D005415:Flavins / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D026901:Membrane Transport Proteins; D011485:Protein Binding; D013050:Spectrometry, Fluorescence
chemical_list: D002352:Carrier Proteins; D005415:Flavins; D026901:Membrane Transport Proteins; C019079:riboflavin-binding protein