Title
Urea Stress: Myo-inositol’s efficacy to counteract destabilization of TIM-β-globin complex by urea is as good as that of the methylamine
Author
Wahiduzzaman, Md. Imtaiyaz Hassan, Asimul Islam, Faizan Ahmad
Year
2019
Journal
International Journal of Biological Macromolecules
Abstract
Do kidney osmolytes counteract the deleterious effects of urea on kidney proteins? To answer this question, we measured guanidinium chloride (GdmCl)-induced denaturation of triose phosphate isomerase-β-globin subunit complex (TIM-β-globin) from sheep kidney in the presence of various concentrations of urea and each of kidney osmolytes (glycine betaine, myo-inositol and sorbitol) alone and in combination at pH 7.5 and 25 °C. Analysis of GdmCl-induced denaturation curve at a given osmolyte (or urea) concentration gave ΔGD0 (Gibbs free energy change in the absence of GdmCl). For a co-solute (CO), a linear plot of ΔΔGD0 (difference between values of ΔGD0 in the presence and absence of CO) against [CO] (molar concentration of CO) yielded m-value (=δΔΔGD0/δ[CO]). m-value for urea (murea) and that for the osmolyte (mosm) were used to determine [urea]:[osmolyte], the molar ratio for the perfect counteraction of the denaturing effect of urea on the protein stability by an osmolyte. For each osmolyte, we tested this ratio for the perfect counteraction. It was observed that glycine betaine and myo-inositol provide perfect counteraction at the predicted molar ratio whereas sorbitol fails to do so. Furthermore, urea and GdmCl induced a two-state denaturation, and both TIM and β-globin have identical thermodynamic stability.
Instrument
J-1500, V-660
Keywords
Circular dichroism, Absorption, Protein structure, Protein denaturation, Secondary structure, Protein folding, Ligand binding, Thermodynamics, Biochemistry