title: The effect of water on the rate of conformational change in protein allostery.
abstract: The influence of solvation on the rate of quaternary structural change is investigated in human hemoglobin, an allosteric protein in which reduced water activity destabilizes the R state relative to T. Nanosecond absorption spectroscopy of the heme Soret band was used to monitor protein relaxation after photodissociation of aqueous HbCO complex under osmotic stress induced by the nonbinding cosolute poly(ethylene glycol) (PEG). Photolysis data were analyzed globally for six exponential time constants and amplitudes as a function of osmotic stress and viscosity. Increases in time constants associated with geminate rebinding, tertiary relaxation, and quaternary relaxation were observed in the presence of PEG, along with a decrease in the fraction of hemes rebinding CO with the slow rate constant characteristic of the T state. An analysis of these results along with those obtained by others for small cosolutes showed that both osmotic stress and solvent viscosity are important determinants of the microscopic R --> T rate constant. The size and direction of the osmotic stress effect suggests that at least nine additional water molecules are required to solvate the allosteric transition state relative to the R-state hydration, implying that the transition state has a greater solvent-exposed area than either end state.
authors.affiliation: Department of Chemistry and Biochemistry, University of California, Santa Cruz, California 95064, USA. goldbeck@chemistry.ucsc.edu
authors.forename: R A
authors.initials: RA
authors.lastname: Goldbeck
authors.forename: S J
authors.initials: SJ
authors.lastname: Paquette
authors.forename: D S
authors.initials: DS
authors.lastname: Kliger
mesh_terms: D000494:Allosteric Regulation / Q000187:drug effects / Q000502:physiology; D001665:Binding Sites / Q000187:drug effects / Q000502:physiology; D002248:Carbon Monoxide / Q000378:metabolism*; D006454:Hemoglobins / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008954:Models, Biological; D009997:Osmotic Pressure; D010782:Photolysis / Q000187:drug effects; D011092:Polyethylene Glycols / Q000494:pharmacology*; D011487:Protein Conformation / Q000187:drug effects; D020836:Protein Structure, Quaternary / Q000187:drug effects / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000187:drug effects / Q000502:physiology; D014867:Water / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D006454:Hemoglobins; D014867:Water; D011092:Polyethylene Glycols; D002248:Carbon Monoxide