We have developed a series of detergent arrays for high-throughput screening of cell-free protein expression of membrane proteins.
The wheat germ-based in vitro translation assay was performed in a 96-well bilayer format. Each detergent was tested at several concentrations ranging from below the critical micelle concentration (CMC) to several-fold higher than the CMC. Effect on translation was monitored by the level of fluorescence of GFP expressed in each condition. Non-ionic and zwitterionic detergents commonly used for membrane protein purification were tested. Each translation series was performed twice. The results demonstrate the diversity of translation compatibility profiles for commonly used detergents. In addition, a series of novel amphiphiles of the CFA and MNG class were also tested to establish a link between the size of the hydrophilic group and translational compatibility. These low CMC compounds are largely compatible with translation even at concentrations several hundred times the CMC. The compounds with the longest hydrophobic tails displayed the longest hydrophobic tails displayed the highest level of compatibility.
Brian G. Fox (email@example.com)
Transmembrane Protein Center
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Last edited:Mon 01 Oct 2012 - 6 years, 6 months ago