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Protein Sci. 2013 Jun;22(6):840-50. doi: 10.1002/pro.2254. Epub 2013 Apr 29.

A multipurpose fusion tag derived from an unstructured and hyperacidic region of the amyloid precursor protein.

Author information

1
Laboratory of Protein Synthesis and Expression, Institute for Protein Research, Osaka University, Suita, Osaka, Japan.

Abstract

Expression and purification of aggregation-prone and disulfide-containing proteins in Escherichia coli remains as a major hurdle for structural and functional analyses of high-value target proteins. Here, we present a novel gene-fusion strategy that greatly simplifies purification and refolding procedure at very low cost using a unique hyperacidic module derived from the human amyloid precursor protein. Fusion with this polypeptide (dubbed FATT for Flag-Acidic-Target Tag) results in near-complete soluble expression of variety of extracellular proteins, which can be directly refolded in the crude bacterial lysate and purified in one-step by anion exchange chromatography. Application of this system enabled preparation of functionally active extracellular enzymes and antibody fragments without the need for condition optimization.

PMID:
23526492
PMCID:
PMC3690722
DOI:
10.1002/pro.2254
[Indexed for MEDLINE]
Free PMC Article
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