Research objectives and content
P-glycoprotein (PGP) is a membrane protein and its overexpression is linked to the occurrence of multidrug resistance (MDR). The molecular mechanism of PGP as well as its three-dimensional structure are unresolved and are both of enormous biochemical interest and clinical importance. The group at the host laboratory has overexpressed and purified both nucleotide-binding domains (NBD) of PGP. Using recombinant NBDs we attempt the following experiments.
1. The crystallisation of both domains to initiate X-ray crystallographic studies. It is generally intended co-crystallise the protein with a mechanistically relevant ligand.
2. The identification of binding sites for flavonoides and steroides which have been reported to act as modulators of MDR culture cells. Using highly purified PGP which is available for studies at the host laboratory we attempt to establish a system to determine the inhibitory capacities of modulators identified in experiments with the recombinant NBDs. Highly purified PGP will be inserted into liposomes to restore optimal activity of drug transport coupled to ATP hydrolysis and the effect of identified modulators ill be monitored.
Training content (objective, benefit and expected impact)
The applicant will receive training in working with membrane proteins in general and will especially learn to apply fluorescence spectroscopy to address biochemical problems.
Links with industry / industrial relevance (22)
Although no formal link is presently settled with the industry, the project is obviously of considerable interest for pharmaceutical and clinical purposes in the area of anticancer chemotherapy. As soon as potent inhibitors of multidrug resistance are identified appropriate contacts will be established for a possible industrial development and clinical application.