PT - JOURNAL ARTICLE AU - Kaoru Fujimoto AU - Akinobu Senoo AU - Satoru Nagatoishi AU - Daisuke Takahashi AU - Tadashi Ueda AU - Kouhei Tsumoto AU - Jose M.M. Caaveiro TI - Lipid nanodiscs facilitate the identification of a fragment compound inhibiting the enzymatic activity of the bacterial membrane protein MsbA AID - 10.1101/2021.06.15.448612 DP - 2021 Jan 01 TA - bioRxiv PG - 2021.06.15.448612 4099 - http://biorxiv.org/content/early/2021/06/16/2021.06.15.448612.short 4100 - http://biorxiv.org/content/early/2021/06/16/2021.06.15.448612.full AB - Membrane proteins are critical elements of numerous therapeutic approaches ranging from cancer to bacterial infections. MsbA is a bacterial membrane protein that has received increasing attention as an antibacterial target for its role in the processing of Lipid A, a key precursor of lipopolysaccharide that is essential for bacterial growth. When employing nanodiscs it is possible to stabilize MsbA by providing a membrane-like environment that enhances its enzymatic activity. Taking advantage of this property we have carried out a fragment screening using the biophysical method of surface plasmon resonance. This approach identified several compounds that bind specifically to MsbA. In particular, one of these fragment molecules not only binds to the target, but also inhibits the ATPase activity of the MsbA protein. The similarity of this fragment to the adenine moiety of ATP points at a route to generate stronger and more potent inhibitors for MsbA and even other proteins of its family of ABC transporters. Collectively, our study reveals biophysical approaches that facilitate the identification of fragment candidates inhibiting the activity of membrane proteins.Competing Interest StatementThe authors have declared no competing interest.E. coliEscherichia coliLPSLipopolysaccharideDTTDithiothreitolDDMn-dodecyl-β-D-maltosideDMPC1,2-dimyristoyl-sn-glycero-3-phosphocholineMSP1Membrane scaffold proteinAMP-PNP5’-adenylimiimidodiphosphateE. coliEscherichia coliSPRSurface plasmon resonance,ITCIsothermal titration calorimetry,TEVTobacco etch virusSECSize exclusion chromatographySEStandard errorDMSODimethyl sulfoxide