My collaborator was successful in purifying soluble rat phosphatidylinositol transfer protein in E. coli when overexpressing GroEL/ES. They estimated ~50% of the protein became soluble. We were able to solve the structure from protein purified using this expression construct. We did not experience copurification problems.
Yoder MD, Thomas LM, Tremblay JM, Oliver RL, Yarbrough LR, Helmkamp GM Jr. (2001) J. Biol. Chem. 276:9246-9252. We have tried it with other proteins, but not as successfully. Marilyn Marilyn D. Yoder Division of Cell Biology & Biophysics School of Biological Sciences University of Missouri-Kansas City 5007 Rockhill Rd. Kansas City, MO 64110 Phone: 816-235-1986 From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Kothe, Michael Sent: Friday, April 22, 2011 10:28 AM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] Using chaperones to boost expression in E. coli Dear ccp4bb, I am curious to hear of examples where the expression of well-behaved protein was achieved by the coexpression of chaperones in E. coli. I know the appropriate strains and vectors exist, but I can't remember hearing of a successful case. I have heard anecdotally of several cases where it was tried without success (including one attempt I made myself). I also heard of concerns that the chaperones might copurify with the (now soluble) protein of interest and are difficult to remove. Thanks, Michael
