title: Approaches for the generation of active papain-like cysteine proteases from inclusion bodies of Escherichia coli.
abstract: Papain-like cysteine proteases are widely expressed, fulfill specific functions in extracellular matrix turnover, antigen presentation and processing events, and may represent viable drug targets for major diseases. In depth and rigorous studies of the potential for these proteins to be targets for drug development require sufficient amounts of protease protein that can be used for both experimental and therapeutic purposes. Escherichia coli was widely used to express papain-like cysteine proteases, but most of those proteases are produced in insoluble inclusion bodies that need solubilizing, refolding, purifying and activating. Refolding is the most critical step in the process of generating active cysteine proteases and the current approaches to refolding include dialysis, dilution and chromatography. Purification is mainly achieved by various column chromatography. Finally, the attained refolded proteases are examined regarding their protease structures and activities.
authors: School of Life Science, South China Normal University, 55# Zhongshan Road West, Tianhe District, Guangzhou, 510631, People's Republic of China.
authors: Chunfang
authors: C
authors: Ling
authors: Junyan
authors: J
authors: Zhang
authors: Deqiu
authors: D
authors: Lin
authors: Ailin
authors: A
authors: Tao
mesh_terms: D002853:Chromatography, Liquid; D057056:Cysteine Proteases / Q000235:genetics / Q000302:isolation & purification*; D004926:Escherichia coli / Q000166:cytology / Q000201:enzymology* / Q000235:genetics; D002479:Inclusion Bodies / Q000201:enzymology*; D058849:Protein Refolding; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification
chemical_list: D011994:Recombinant Proteins; D057056:Cysteine Proteases