Journal
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
Volume 75, Issue 16, Pages 5356-5362Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/AEM.00691-09
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Funding
- NIGMS NIH HHS [R01 GM085449, U54-GM074958, 1R01GM085449, U54 GM074958] Funding Source: Medline
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Protein S is a major spore coat protein of Myxococcus xanthus, consisting of two homologous domains, the N-terminal domain (NTD) and the C-terminal domain, both of which contain a Ca2+-binding site. Protein S tightly binds to myxospores in a Ca2+-dependent manner. Here, we constructed a novel expression vector, pCold-PST, encoding two tandem repeat NTDs (PrS2). By using this vector, a number of human proteins that were expressed at low levels or in insoluble forms by a pET vector were expressed not only at high levels but also in soluble forms. We also demonstrated that an Escherichia coli protein tagged with PrS2 fully retained its function, indicating that it is folded independently from the tag. This technology not only allows simple, one-step protein purification using myxospores, but can also be used for the identification of proteins interacting with a protein of interest and will prove immensely useful for structural studies of proteins which are difficult to produce or are insoluble.
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