Influence of protein aggregation and macromolecular crowing on the dynamics and function of protiens respectively

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The phenomenon of macromolecular crowding is ubiquitous in cells. It is reported to affect equilibria and kinetics of processes taking place in cells and is also hypothesized to affect enzyme catalyzed reactions. A part of this doctoral work was directed towards ascertaining the effect of crowding on an enzyme catalyzed reaction in an environment crowded with synthetic crowding agents viz., dextran (15-500 kDa ) and Ficoll (70 and 400 kDa). The hydrolysis of p-nitrophenyl phosphate by alkaline phosphatase was used as the model reaction for the purpose.We observed that macromolecular crowding affects the catalytic rate of hydrolysis of p-nitro phenyl phosphate in a size dependent manner. In the presence of increasing concentrations of 15-70 kDa dextrans, the rates were observed to be partly transition rate limited and partly diffusion controlled. However, in the presence of the larger ...
Supervisor: R. Swaminathan