Frank Bernhard, April 2016
Frank Bernhard will visit Caltech on 22 April 2016 (Fri). Please sign up below if you would like to meet with him:
Schedule
22 April 2016 (Friday):
- 9-11 am: Caltech/Amgen project discussion
- 11 am: Talk (room TBD)
- 12 pm: Lunch with Richard, Ching, Amit, Shaobin
- 1:30 pm: Yong
- 2:00 pm: Open
- 2:45 pm: Open
- 3:30 pm: Open
- 4:15 pm: Open
- 5:00 pm: Done for the day
Talk abstract
Frank Bernhard
Institute of Biophysical Chemistry
Center for Biomolecular Magnetic Resonance
INSTRUCT Core Center for Cell-Free Expression
J.W. Goethe-University, Frankfurt-am-Main 60438, Germany
Cell-free expression technologies represent a continuously growing central platform in the emerging field of synthetic biology. Core applications are the synthesis of difficult biomolecules that cannot be obtain in cellular production systems due to toxic effects, absence of proper folding environments or other limitations. Bacterial lysates of certain E. coli strains combine high production efficiencies with considerable tolerance for a wide range of supplied compounds such as stabilizer, cofactors or ligands. Cell-free expression environments can therefore be adjusted according to the individual requirements of synthesized target proteins. We have analyzed the proteome background of lysates processed for high expression efficiency by detailed proteomics studies. Effects of lysate modifications on proteome composition and quality of synthesized targets was further analyzed. In addition, we exemplify the screening of beneficial additives such as chemical chaperones.
Membrane proteins are key targets of current drug and antibody development and often difficult to synthesize in sufficient amounts and quality by using conventional cell-based production systems. Cell-free systems reduce complexity of membrane protein production to the central translation process and basic production protocols can be standardized. The open nature of cell-free systems furthermore allows to create customized and well-defined hydrophobic environments composed out of detergents, lipids or amphipols. Cell-free expression therefore completely re-designs natural targeting and folding pathways of membrane proteins. By using preformed membrane-like particles such as nanodiscs, even detergent sensitive membrane proteins can be synthesized and subsequently characterized at the molecular level. We exemplify the expression tuning of a variety of membrane proteins comprising G-protein coupled receptors, transporters or large complexes.
