Kumar, RajeshKumar, RajeshSharma, DeepakGarg, MansiKumar, VinayAgarwal, Mukesh Chand2019-03-222024-08-132019-03-222024-08-132018Kumar R., Kumar R., Sharma D. et.al. (2018) Macromolecular crowding-induced molten globule states of the alkali pH-denatured proteins1570963910.1016/j.bbapap.2018.08.012https://kr.cup.edu.in/handle/32116/2035Structural and molecular properties extracted from circular dichroism (CD), tryptophan fluorescence and 1-anilino-8-napthalene sulfonate (ANS) binding experiments suggest that the high concentration of synthetic crowding agents (dextran 40, dextran 70 and ficoll 70) stabilizes and refolds the base-denatured ferricytochrome c (Ferricyt c) and lysozyme (Lyz) at pH 12.9 (±0.1) to molten globule (MG) states (C B -states). These results further revealed that the C B -states resemble the generic properties of MG-states. Thermodynamic analysis of thermal denaturation curves of base-denatured Ferricyt c and Lyz at pH 12.9 (±0.1) under variable concentrations of crowding agents (dextran 40, dextran 70 and ficoll 70) revealed that the crowder presence increases the thermal stability of base-denatured proteins and also prevents the cold denaturation of Ferricyt c. The results further showed that the nature, size and shape of crowder influence the crowding-mediated increase in secondary structure stabilization and thermal stability of base-denatured Ferricyt c and Lyz. Analysis of kinetic and thermodynamic parameters measured for CO association reaction of alkaline ferrocytochrome c (Ferrocyt c) at pH 12.9 (±0.1) under variable concentrations of crowding agents (dextran 40, dextran 70 and ficoll 70) revealed that the crowder presence reduces the level of structural fluctuation of M80-containing ?-loop that control CO association to alkaline Ferrocyt c. - 2018 Elsevier B.V.en-USCold denaturationCrowding agentsEnthalpy-entropy plotMolten-globule stateThermal stabilityArticlehttps://www.sciencedirect.com/science/article/pii/S157096391830147X