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Purification of Hsp104, a Protein Disaggregase

Article DOI: 10.3791/3190-v • 07:18 min • September 30th, 2011
September 30th, 2011 •
1, 1, 1
1Department of Biochemistry and Biophysics, University of Pennsylvania

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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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Purification Hsp104 Protein Disaggregase Hexameric AAA+ Protein ATP Hydrolysis Protein Disaggregation Renaturation Disordered Aggregates Heat Shock Cross-beta Amyloid Fibrils Prions Phenotypic Variation Preamyloid Oligomers E. Coli Orthologue ClpB Hsp104 Orthologues Animal Cells Therapeutic Agent Neurodegenerative Diseases Misfolding Of Proteins Toxic Preamyloid Oligomers Amyloid Fibrils
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