Expression and purification of the recombinant membrane protein YidC: A case studyfor increased solubility and stability

Expression and purification of the recombinant membrane protein YidC : A case studyfor increased solubility and stability

Molina, Daniel Martinez (author): Stockholms universitet,Institutionen för biokemi och biofysik

Niegowski, Damian (author): Stockholms universitet,Institutionen för biokemi och biofysik

(creator_code:org_t)

Elsevier BV, 2008
2008
English.
In: Protein Expression and Purification. - : Elsevier BV. - 1046-5928 .- 1096-0279. ; 62:1, s. 49-52

Related links:: https://urn.kb.se/re...; show more...; https://doi.org/10.1...; https://urn.kb.se/re...; http://kipublication...; show less...

Abstract Subject headings

YidC is an inner membrane protein from Escherichia coli and is an essential component in insertion, trans- location 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 lg 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 con- centrations and that was stable for weeks at +4 C. The isolated protein is thus well suited for structural studies.

MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)
MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)
TEKNIK OCH TEKNOLOGIER -- Medicinteknik -- Medicinsk laboratorie- och mätteknik (hsv//swe)
ENGINEERING AND TECHNOLOGY -- Medical Engineering -- Medical Laboratory and Measurements Technologies (hsv//eng)

By the author/editor: Molina, Daniel M ...; Lundbäck, Anna-K ...; Niegowski, Damia ...; Eshaghi, Said

About the subject

MEDICAL AND HEALTH SCIENCES: MEDICAL AND HEAL ...; and Medical Biotechn ...; and Medical Biotechn ...

ENGINEERING AND TECHNOLOGY: ENGINEERING AND ...; and Medical Engineer ...; and Medical Laborato ...

By the university: Stockholm University; Royal Institute of Technology; Karolinska Institutet

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