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

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dc.contributor.author PANDE, VANI en_US
dc.contributor.author PANANGHAT, GAYATHRI,  en_US
dc.date.accessioned 2024-10-29T06:44:39Z
dc.date.available 2024-10-29T06:44:39Z
dc.date.issued 2024-10 en_US
dc.identifier.citation Bio-protocol, 14(20). en_US
dc.identifier.issn 2331-8325 en_US
dc.identifier.uri https://doi.org/10.21769/BioProtoc.5086 en_US
dc.identifier.uri http://dr.iiserpune.ac.in:8080/xmlui/handle/123456789/9142
dc.description.abstract 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 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 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. en_US
dc.language.iso en en_US
dc.publisher Bio-protocol LLC en_US
dc.subject MreB  en_US
dc.subject Cytoskeleton proteins en_US
dc.subject ADP en_US
dc.subject Thermal shift assay Affinity chromatography en_US
dc.subject Size exclusion chromatography en_US
dc.subject Crystallization en_US
dc.subject X-Ray adsorption spectroscopy en_US
dc.subject 2024 en_US
dc.subject 2024-OCT-WEEK2 en_US
dc.subject TOC-OCT-2024  en_US
dc.title Improving Stability of Spiroplasma citri MreB5 Through Purification Optimization and Structural Insights en_US
dc.type Article en_US
dc.contributor.department Dept. of Biology en_US
dc.identifier.sourcetitle Bio-protocol en_US
dc.publication.originofpublisher Foreign en_US


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