RT Journal Article SR Electronic T1 Production and purification of human Hsp90β in Escherichia coli JF bioRxiv FD Cold Spring Harbor Laboratory SP 121996 DO 10.1101/121996 A1 Martina Radli A1 Dmitry B. Veprintsev A1 Stefan G. D. Rüdiger YR 2017 UL http://biorxiv.org/content/early/2017/03/30/121996.abstract AB The molecular chaperone Hsp90 is an essential member of the cellular proteostasis system. It plays an important role in the stabilisation and activation of a large number of client proteins and is involved in fatal disease processes e.g. Alzheimer disease, cancer and cystic fibrosis. This makes Hsp90 a crucial protein to study. Mechanistic studies require large amounts of protein but the production and purification of recombinant human Hsp90 in E. coli is challenging and laborious. Here we identified conditions that influence Hsp90 production and optimised a fast and efficient purification protocol. We found that the nutrient value of the culturing medium and the length of induction had significant effect on Hsp90 production in Escherichia coli. Our fast, single-day purification protocol resulted in a stable, well-folded and pure sample that was resistant to degradation in a reproducible manner. We anticipate that our results provide a useful tool to produce higher amount of pure, well-folded and stable recombinant human Hsp90β in Escherichia coli in an efficient way.