Improving Stability of Spiroplasma citri MreB5 Through Purification Optimization and Structural Insights

Improving Stability of Spiroplasma citri MreB5 Through Purification Optimization and Structural Insights

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MreB is a prokaryotic actin homolog.It is essential for cell shape in the majority of rod-shaped cell-walled bacteria.Structural and functional characterization of MreB protein is important Keepsakes to understand the mechanism of ATP-dependent filament dynamics and membrane interaction.In vitro studies on MreBs have been limited due to the difficulty in purifying the homogenous monomeric protein.We have purified MreB from the cell-wall-less bacteria Spiroplasma citri, ScMreB5, using heterologous expression in Escherichia coli.

This protocol provides a detailed description of purification condition optimization that led us to obtain high Bike Parts - BMX - Wheel Rims concentrations of stable ScMreB5.Additionally, we have provided a protocol for detecting the presence of monovalent ions in the ScMreB5 AMP-PNP-bound crystal structure.This protocol can be used to obtain a high yield of ScMreB5 for carrying out biochemical and reconstitution studies.The strategies used for ScMreB5 show how optimizing buffer components can enhance the yield and stability of purified protein.