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.
| Original language | English |
|---|---|
| Journal | FEBS Letters |
| Volume | 584 |
| Issue number | 4 |
| Pages (from-to) | 669-674 |
| Number of pages | 6 |
| ISSN | 0014-5793 |
| DOIs | |
| Publication status | Published - 02.2010 |
Funding
We thank Frederice Gries for technical assistance and Gunter Stier for the gift of pETM-Z. We would like to express our appreciation to the Kaufmann lab (Max Planck Institute for Infectious Biology, Berlin, Germany) for providing the initial pSD26 vector. We would also like to thank the Mandelkow Laboratory (Max Planck Unit for Structural Molecular Biology, Hamburg, Germany) for electron microscope access and Gerard Drewes (Cellzome, Heidelberg) and Matthias Ehebauer for critical reading of the manuscript. This work was supported by the EC Grant ScrIn-Silico ( LSHP-CT-012127 ) to M.W., by the BMBF Grant “X-MTB” ( 0312992A ) to M.W., and by a grant within the BMBF programme Pathogenomik Plus ( PTJ-BIO 0313801L ) to M.W.
