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    Factors defining the effects of macromolecular crowding on dynamics and thermodynamic stability of heme proteins in-vitro,
    (Elsevier, 2018) Kumar, Rajesh; Sharma, Deepak; Kumar, Vinay; Kumar, Rajesh
    The role of crowding agents on structure and activities of heme proteins has been established. Analysis of kinetic and thermodynamic parameters measured for CO-dissociation reaction of natively-folded carbonmonoxycytochrome c (NCO) and carbonmonoxymyoglobin (MbCO) at different [GdnHCl] or [Urea] in the presence of crowding agents (dextran 40, dextran 70 and ficoll 70) demonstrate that (i) at low denaturant concentrations, crowder presence enhances the denaturant-mediated restricted dynamics of NCO and MbCO, and (ii) at higher denaturant concentrations, large scale unfolding-fluctuations dominate the dynamics and inclusion of crowder counteracts the structural-fluctuations causing the unfolding of proteins. Thermodynamic analysis of thermal and urea-unfolding curves of cytochrome c (Cyt c) and myoglobin (Mb) measured at different [GdnHCl] in presence of crowding agents reveals that crowder presence counterbalances and strengthens the destabilizing action of GdnHCl on stability of Cyt c and Mb, respectively. This study further demonstrates that the size, shape and concentration of crowding agent modulate the effect of crowder on denaturant-mediated dynamics and thermodynamic stability of heme proteins.
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    Optimization of salt concentration and explanation of two peak percolation in blend solid polymer nanocomposite films
    (Springer New York LLC, 2018) Arya, A.; Sharma, A.L.
    The present paper is focused toward the preparation of the flexible and free-standing blend solid polymer electrolyte films based on PEO-PVP complexed with NaPF6 by the solution cast technique. The structural/morphological features of the synthesized polymer nanocomposite films have been investigated in detail using X-ray diffraction, Fourier transform infra-red spectroscopy, Field emission scanning electron microscope, and Atomic force microscopy techniques. The film PEO-PVP + NaPF6 ((Formula presented.)8) exhibits highest ionic conductivity ~ 5.92 ? 10?6 S cm?1 at 40 ?C and ~ 2.46 ? 10?4 S cm?1 at 100 ?C. The temperature-dependent conductivity shows an Arrhenius type behavior and activation energy decreases with the addition of salt. The high temperature (100 ?C) conductivity monitoring is done for the optimized PEO-PVP + NaPF6 ((Formula presented.)8) highly conductive system and the conductivity is still maintained stable up to 160 h (approx. 7 days). The thermal transitions parameters were measured by the differential scanning calorimetry (DSC) measurements. The prepared polymer electrolyte film displays the smoother surface on addition of salt and a thermal stability up to 300 ?C. The ion transference number (tion) for the highest conducting sample is found to be 0.997 and evidence that the present system is ion dominating with negligible electron contribution. Both linear sweep voltammetry and cyclic voltammetry supports the use of prepared polymer electrolyte with long-term cycle stability and thermal stability for the solid-state sodium ion batteries. Finally, a two peak percolation mechanism has been proposed on the basis of experimental findings. [Figure not available: see fulltext.] ? 2018 Springer-Verlag GmbH Germany, part of Springer Nature
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    Analysis of the pH-dependent thermodynamic stability, local motions, and microsecond folding kinetics of carbonmonoxycytochrome c
    (Academic Press Inc., 2016) Kumar, Rajesh
    This paper analyzes the effect of pH on thermodynamic stability, low-frequency local motions and microsecond folding kinetics of carbonmonoxycytochrome c (Cyt-CO) all across the alkaline pH-unfolding transition of protein. Thermodynamic analysis of urea-induced unfolding transitions of Cyt-CO measured between pH 6 and pH 11.9 reveals that Cyt-CO is maximally stable at pH?9.5. Dilution of unfolded Cyt-CO into refolding medium forms a native-like compact state (NCO-state), where Fe2+?CO interaction persists. Kinetic and thermodynamic parameters measured for slow thermally-driven CO dissociation (NCO?N+CO) and association (N+CO?NCO) reactions between pH 6.5 and pH 13 reveal that the thermal-motions of M80-containing ?-loop are decreased in subdenaturing limit of alkaline pH. Laser photolysis of Fe2+-CO bond in NCO-state triggers the microsecond folding (NCO?N). The microsecond kinetics measured all across the alkaline pH-unfolding transition of Cyt-CO produce rate rollover in the refolding limb of chevron plot, which suggests a glass transition of NCO en route to N. Between pH 7 and pH 11.9, the natural logarithm of the microsecond folding rate varies by?<?1.5 units while the natural logarithm of apparent equilibrium constant varies by 11.8 units. This finding indicates that the pH-dependent ionic-interactions greatly affect the global stability of protein but have very small effect on folding kinetics. ? 2016 Elsevier Inc.