Expression and purification of the recombinant membrane protein YidC: A case study for increased stability and solubility
2008 (English)In: Protein Expression and Purification, ISSN 1046-5928, E-ISSN 1096-0279, Vol. 62, no 1, 49-52 p.Article in journal (Refereed) Published
YidC is an inner membrane protein from Escherichia coli and is an essential component in insertion, translocation and assembly of membrane proteins in the membranes. Previous purification attempts resulted in heavy aggregates and precipitated protein at later stages of purification. Here we present a rapid and straightforward stability screening strategy based on gel filtration chromatography, which requires as little as 10 mu g of protein and takes less than 15 min to perform. With this technique, we could rapidly screen several buffers in order to identify an optimum condition that stabilizes purified YidC. After optimization we could obtain several milligrams of purified YidC that could be easily prepared at high concentrations and that was stable for weeks at +4 degrees C. The isolated protein is thus well suited for structural studies.
Place, publisher, year, edition, pages
2008. Vol. 62, no 1, 49-52 p.
Membrane protein insertion system, Stability optimization, Analytical size exclusion chromatography, Gel filtration, Buffer screen
Medical Laboratory and Measurements Technologies
IdentifiersURN: urn:nbn:se:kth:diva-36398DOI: 10.1016/j.pep.2008.05.011ISI: 000260061900008ScopusID: 2-s2.0-52949101714OAI: oai:DiVA.org:kth-36398DiVA: diva2:430672
QC 201107122011-07-122011-07-122011-07-12Bibliographically approved