Spadaccini, Roberta, Hundeep Kaur, Johanna Becker-Baldus, and Clemens Glaubitz. “The Effect of Drug Binding on Specific Sites in Transmembrane Helices 4 and 6 of the ABC Exporter MsbA Studied by DNP-Enhanced Solid-State NMR.” Biochimica et Biophysica Acta (BBA) – Biomembranes 1860, no. 4 (April 2018): 833–40.
MsbA, a homodimeric ABC exporter, translocates its native substrate lipid A as well as a range of smaller, amphiphilic substrates across the membrane. Magic angle sample spinning (MAS) NMR, in combination with dynamic nuclear polarization (DNP) for signal enhancement, has been used to probe two speciﬁc sites in transmembrane helices 4 and 6 of full length MsbA embedded in lipid bilayers. Signiﬁcant chemical shift changes in both sites were observed in the vanadate-trapped state compared to apo state MsbA. The reduced spectral line width indicates a more conﬁned conformational space upon trapping. In the presence of substrates Hoechst 33342 and daunorubicin, further chemical shift changes and line shape alterations mainly in TM6 in the vanadate trapped state were detected. These data illustrate the conformational response of MsbA towards the presence of drugs during the catalytic cycle.