title: Effects of alcohols on the stability and low-frequency local motions that control the slow changes in structural dynamics of ferrocytochrome c.
abstract: To determine the effects of alcohols on the low-frequency local motions that control slow changes in structural dynamics of native-like compact states of proteins, we have studied the effects of alcohols on structural fluctuation of M80-containing Ω-loop by measuring the rate of thermally driven CO dissociation from a natively folded carbonmonoxycytochrome c under varying concentrations of alcohols (methanol, ethanol, 1-propanol, 2-propanol, 3°-butanol, 2,2,2-trifluoroethanol). As alcohol is increased, the rate coefficient of CO dissociation (k(diss)) first decreases in subdenaturing region and then increases on going from subdenaturing to denaturing milieu. This decrease in k(diss) is more for 2,2,2-trifluroethanol and 1-propanol and least for methanol, indicating that the first phase of motional constraint is due to the hydrophobicity of alcohols and intramolecular protein cross-linking effect of alcohols, which results in conformational entropy loss of protein. The thermal denaturation midpoint for ferrocytochrome c decreases with increase in alcohol, indicating that alcohol decrease the global stability of protein. The stabilization free energy (ΔΔG) in alcohols' solution was calculated from the slope of the Wyman-Tanford plot and water activity. The m-values obtained from the slope of ΔΔG versus alcohols plot were found to be more negative for longer and linear chain alcohols, indicating destabilization of proteins by alcohols through disturbance of hydrophobic interactions and hydrogen bonding.
authors.affiliation: School of Chemistry and Biochemistry, Thapar University, Patiala 147004; and Council of Scientific and Industrial Research-Institute of Microbial Technology, Sector 39A, Chandigarh, India.
authors.forename: Rishu
authors.initials: R
authors.lastname: Jain
authors.forename: Deepak
authors.initials: D
authors.lastname: Sharma
authors.forename: Rajesh
authors.initials: R
authors.lastname: Kumar
mesh_terms: D000438:Alcohols / Q000737:chemistry*; D000818:Animals; D002942:Circular Dichroism; D045304:Cytochromes c / Q000737:chemistry*; D004795:Enzyme Stability; D006736:Horses; D006860:Hydrogen Bonding; D057927:Hydrophobic and Hydrophilic Interactions; D008958:Models, Molecular; D011489:Protein Denaturation; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D013816:Thermodynamics; D014867:Water / Q000737:chemistry
chemical_list: D000438:Alcohols; D014867:Water; D045304:Cytochromes c
keywords: WT plot; constrained dynamics; m-value; stabilization free energy; van’t Hoff enthalpy; water activity