Open Access Research article

Overproduced Brucella abortus PdhS-mCherry forms soluble aggregates in Escherichia coli, partially associating with mobile foci of IbpA-YFP

Charles Van der Henst1, Caroline Charlier2, Michaël Deghelt1, Johan Wouters2, Jean-Yves Matroule1, Jean-Jacques Letesson1 and Xavier De Bolle1*

Author Affiliations

1 Molecular Biology Research Unit (URBM), University of Namur (FUNDP), 61 rue de Bruxelles, 5000 Namur, Belgium

2 Theoretical and Structural Physical Chemistry Unit (UCPTS), University of Namur (FUNDP), 61 rue de Bruxelles, 5000 Namur, Belgium

For all author emails, please log on.

BMC Microbiology 2010, 10:248  doi:10.1186/1471-2180-10-248

Published: 28 September 2010



When heterologous recombinant proteins are produced in Escherichia coli, they often precipitate to form insoluble aggregates of unfolded polypeptides called inclusion bodies. These structures are associated with chaperones like IbpA. However, there are reported cases of "non-classical" inclusion bodies in which proteins are soluble, folded and active.


We report that the Brucella abortus PdhS histidine kinase fused to the mCherry fluorescent protein forms intermediate aggregates resembling "non-classical" inclusion bodies when overproduced in E. coli, before forming "classical" inclusion bodies. The intermediate aggregates of PdhS-mCherry are characterized by the solubility of PdhS-mCherry, its ability to specifically recruit known partners fused to YFP, suggesting that PdhS is folded in these conditions, and the quick elimination (in less than 10 min) of these structures when bacterial cells are placed on fresh rich medium. Moreover, soluble PdhS-mCherry foci do not systematically colocalize with IpbA-YFP, a marker of inclusion bodies. Instead, time-lapse experiments show that IbpA-YFP exhibits rapid pole-to-pole shuttling, until it partially colocalizes with PdhS-mCherry aggregates.


The data reported here suggest that, in E. coli, recombinant proteins like PdhS-mCherry may transit through a soluble and folded state, resembling previously reported "non-classical" inclusion bodies, before forming "classical" inclusion bodies. The dynamic localization of IbpA-YFP foci suggests that the IbpA chaperone could scan the E. coli cell to find its substrates.