Figure 1
Comparison of Apo and Cargo-loaded Bla g 1. 31P-NMR spectra (a) of Bla g 1 purified directly from E. coli (top), and DSPC-loaded Bla g 1 purified using reverse-phase HPLC followed by the annealing protocol (bottom). The DSPC standard curve used to estimate Bla g 1 binding stoichiometry from 31P-NMR intensities is shown in (b). CD spectra (c) obtained for Bla g 1 purified using reverse-phase HPLC coupled with annealing in the absence of cargo (Black). Spectra of Bla g 1 loaded with nMix, a mixture of palmitate, oleate, and stearate fatty acids which mimics the cargo identified from Bla g 1 obtained from its natural allergen (Red), or 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC) phospholipid (Orange) cargo are shown for comparison. Representative gel-filtration elution profiles (d) for Apo (Black) and nMix-loaded (Red) Bla g 1 generated through the annealing process, and Bla g 1 purified from E. coli without annealing (Green).
