Figure 2: Verifying lipid removal and loading of Bla g 1.
A) 31P-NMR spectra of Apo- (black) or DSPC-loaded Bla g 1 (red) prepared using the annealing protocol described in this work demonstrating the complete removal of lipids in the former, and the homogeneous loading of phosphatidylcholine (PC) lipids achieved in the latter. In contrast, Bla g 1 purified from recombinant E. coli without lipid stripping and annealing (ecBla g 1) shows a heterogeneous mixture of endogenous phosphatidylethanolamine (PE) and phosphatidylglycerol (PG) lipids when analyzed using this method (B). A representative standard curve obtained from DSPC reference samples of known concentrations is shown in (C), from which the Bla g 1 binding stoichiometry can be obtained. Figures adapted from Foo et al. (2019) and presented under the Creative Commons CC BY License.18