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September 30, 2011
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Here, we describe a protocol for the purification of highly active Hsp104, a hexameric AAA+ protein from yeast, which couples ATP hydrolysis to protein disaggregation. This scheme exploits a His6-tagged construct for affinity purification from E. coli followed by anion-exchange chromatography, His6-tag removal with TEV protease, and size-exclusion chromatography.
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Sweeny, E. A., DeSantis, M. E., Shorter, J. Purification of Hsp104, a Protein Disaggregase. J. Vis. Exp. (55), e3190, doi:10.3791/3190 (2011).
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