Significant enhanced expression and solubility of human proteins in Escherichia coli by fusion with protein S from Myxococcus xanthus.
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 Ca(2+)-binding site. Protein S tightly binds to myxospores in a Ca(2+)-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.
Kobayashi
Hiroshi
H
Department of Biochemistry, Robert Wood Johnson Medical School, Piscataway, NJ 08854, USA.
Yoshida
Takeshi
T
Inouye
Masayori
M
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005821:Genetic Techniques; D005822:Genetic Vectors; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017510:Protein Folding; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D012995:Solubility; D015854:Up-Regulation
D001426:Bacterial Proteins; D011506:Proteins; D011993:Recombinant Fusion Proteins; C024278:protein S, Myxococcus xanthus