Please use this identifier to cite or link to this item: http://dr.iiserpune.ac.in:8080/xmlui/handle/123456789/9142
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dc.contributor.authorPANDE, VANIen_US
dc.contributor.authorPANANGHAT, GAYATHRI, en_US
dc.date.accessioned2024-10-29T06:44:39Z-
dc.date.available2024-10-29T06:44:39Z-
dc.date.issued2024-10en_US
dc.identifier.citationBio-protocol, 14(20).en_US
dc.identifier.issn2331-8325en_US
dc.identifier.urihttps://doi.org/10.21769/BioProtoc.5086en_US
dc.identifier.urihttp://dr.iiserpune.ac.in:8080/xmlui/handle/123456789/9142-
dc.description.abstractMreB 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.isoenen_US
dc.publisherBio-protocol LLCen_US
dc.subjectMreB en_US
dc.subjectCytoskeleton proteinsen_US
dc.subjectADPen_US
dc.subjectThermal shift assay Affinity chromatographyen_US
dc.subjectSize exclusion chromatographyen_US
dc.subjectCrystallizationen_US
dc.subjectX-Ray adsorption spectroscopyen_US
dc.subject2024en_US
dc.subject2024-OCT-WEEK2en_US
dc.subjectTOC-OCT-2024 en_US
dc.titleImproving Stability of Spiroplasma citri MreB5 Through Purification Optimization and Structural Insightsen_US
dc.typeArticleen_US
dc.contributor.departmentDept. of Biologyen_US
dc.identifier.sourcetitleBio-protocolen_US
dc.publication.originofpublisherForeignen_US
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