- Oral Presentation
- Open Access
Protein aggregation into bacterial inclusion bodies is a specific kinetically driven process
Microbial Cell Factories volume 5, Article number: S9 (2006)
Bacterial inclusion bodies are major bottlenecks in protein production and are hampering the development of top priority research areas such as structural genomics. Inclusion body formation was formerly considered to occur via non-specific association of hydrophobic surfaces in folding intermediates rendering biologically inactive protein. Increasing evidence, however, indicates that protein aggregation in bacteria is a rather specific event which might result in active inclusion bodies [1, 2].
Here, first we have used fluorescence resonance energy transfer and microscopy to investigate the degree to which unrelated proteins expressed in the same cells coaggregate with one another. Our data reveal that in bacteria, protein aggregation is a specific event even among highly aggregation-prone polypeptides expressed at high levels.
Second, we have investigated the effect of a large set of single-point mutants of one of these proteins on its specific activity once deposited in inclusion bodies. We find that the activity of such aggregates significantly correlates with the predicted aggregation rates for each mutant.
Overall the data in this study confirms that in vivo protein aggregation depends on molecular recognition and suggests that rationally tuning the kinetic competition between folding and aggregation might result in highly active, inclusion bodies. The exploration of this technology during recombinant protein production would have a significant biotechnological value.
Carrio M, Gonzalez-Montalban N, Vera A, Villaverde A, Ventura S: Amyloid-like properties of bacterial inclusion bodies. J Mol Biol. 2005, 347: 1025-1037. 10.1016/j.jmb.2005.02.030.
Garcia-Fruitos E, Gonzalez-Montalban N, Morell M, Vera A, Ferraz RM, Aris A, Ventura S, Villaverde A: Aggregation as bacterial inclusion bodies does not imply inactivation of enzymes and fluorescent proteins. Microb Cell Fact. 2005, 4: 27- 10.1186/1475-2859-4-27.
About this article
Cite this article
de Groot, N.S., Morell, M., Vendrell, J. et al. Protein aggregation into bacterial inclusion bodies is a specific kinetically driven process. Microb Cell Fact 5, S9 (2006) doi:10.1186/1475-2859-5-S1-S9
- Inclusion Body
- Protein Aggregation
- Fluorescence Resonance Energy Transfer
- Molecular Recognition
- Resonance Energy Transfer