Structural determination of the key membrane protein in peptidoglycan synthesis, MraY, in the presence of inhibitors
EMSL Project ID
51578
Abstract
In the continuous race against bacterial infections, the development of new drugs becomes essential to find new ways to fight multidrug resistant bacteria. MraY catalyzes the first membrane step in the biosynthesis of peptidoglycan precursor lipid I and is considered a promising new clinical target for the development of antibiotics. Despite recent advances in MraY structure, so far, the key structure bound to a substrate remains to be solved. We are working towards the structural characterization of MraY with different inhibitors and a substrate analog. In addition, some recent studies point to a critical role of lipids in MraY dimerization and proper functioning. Despite the proposed important role, no structure of MraY in a lipidic environment has been obtained. We are using MraY reconstituted in nanodisc and single particle analysis to elucidate MraY in a native-like environment. We developed a purification protocol that allow us to purify the sample at a yield of 200 µg per liter of culture in a homodisperse stable peak. Our preliminary data collection shows well-defined particles with 2D classes showing different nanodisc views. Due to our limited availability of time in our CryoEM facility, we are requesting screening and collection time to optimize the sample and acquire data.
Project Details
Start Date
2020-07-15
End Date
2021-03-17
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members