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

• 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.

Ämnesord

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)

Nyckelord

Membrane proteins insertion system

stability optimization

analytical size exclusion chromatography

gel filtration

buffer screen

Biochemistry

Biokemi

Biogeochemistry

biogeokemi

Medical engineering

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