Optimization of membrane protein production

To other research topics:

Cellular osmoregulation

Bacterial vitamin transport

Amino acid and peptide transport

Yeast Organellar transport

Pore forming peptides

Optimization of membrane protein production (Prof. Dr. B. Poolman, Prof. Dr. D.J. Slotboom) 

• Optimization of gene expression and identification of bottlenecks in protein production. Lactococcus lactis and Escherichia coli are used as hosts for the overexpression of membrane proteins.^1,2 A high-throughput method for cloning in E.coli and L. lactis is available. Expression screening by GFP-tagging of proteins is shown in the picture.
• Hydrodynamic analysis of proteins: analytical ultracentrifugation and multi-angle light scattering-coupled to size-exclusion chromatography are used to determine oligomeric state and monodispersity of membrane proteins.
• Crystallography: in collaboration with the group of Andy-Mark Thunnissen and Bauke Dijkstra, X-ray crystallography is used to determine high-resolution structures of membrane proteins.

1. Kunji, E.R.S., Slotboom, D.-J., and Poolman, B. (2003) Lactococcus lactis as host for overproduction of functional membrane proteins. Biochim. Biophys. Acta, 1610, 97-108.
2. Geertsma, E.R., and Poolman, B. (2007) High-throughput cloning and expression in recalcitrant bacteria. Nature Meth., 4, 705-707.