| | Stoichiometric protein complex formation and over-expression using the prokaryotic native operon structureEdited by Gianni Cesareni Received 20 July 2009; received in revised form 22 December 2009; accepted 22 December 2009. published online 18 January 2010. Abstract In prokaryotes, operon encoded proteins often form protein–protein complexes. Here, we show that the native structure of operons can be used to efficiently overexpress protein complexes. This study focuses on operons from mycobacteria and the use of Mycobacterium smegmatis as an expression host. We demonstrate robust and correct stoichiometric expression of dimers to higher oligomers. The expression efficacy was found to be largely independent of the intergenic distances. The strategy was successfully extended to express mycobacterial protein complexes in Escherichia coli, showing that the operon structure of gram-positive bacteria is also functional in gram-negative bacteria. The presented strategy could become a general tool for the expression of large quantities of pure prokaryotic protein complexes for biochemical and structural studies. European Molecular Biology Laboratory (EMBL), Hamburg Outstation, c/o DESY, Notkestrasse 85, 22607 Hamburg, Germany  Corresponding author. Fax: +49 4089902149.
PII: S0014-5793(10)00042-6 doi:10.1016/j.febslet.2009.12.057 © 2010 Federation of European Biochemical Societies | |
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ABSTRACT
