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

Autor:	Yasushi Saheki, Sanae Tabata, Kei Suzuki, Takeshi Sangawa, Keiji Tanaka, Junichi Takagi
Rok vydání:	2013
Předmět:	Cloning chemistry.chemical_classification Fusion Extracellular proteins biology medicine.disease_cause Biochemistry Enzyme chemistry medicine Amyloid precursor protein biology.protein Extracellular Molecular Biology Escherichia coli Peptide sequence
Zdroj:	Protein Science. 22:840-850
ISSN:	0961-8368
DOI:	10.1002/pro.2254
Popis:	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.
Databáze:	OpenAIRE
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