Supplemental Material Figure legends
S1: Initial characterization of A1LS and JR2LS.
A) 1H-15N HSQC of A1LS before and
after redox buffer. Magenta contours show purified A1LS in PBS
buffer. The blue contours show A1LS following incubation with reduced
and oxidized glutathione (2 and 0.5 mM respectively). New peaks
following treatment are labeled. B) TALOS secondary structure prediction
for A1LS based on backbone and side-chain chemical shifts. The two
helical regions of the predicted α-hairpin fold are clearly
identifiable. C) 1H-15N-NMR spectrum
of FL JR2 LS displaying numerous well-dispersed peaks consistent with an
alpha-helical protein. D) Circular dichroism spectra of indicated
hairpinin and the table below showing the percentage of a-helices,
b-sheets and random coils within each.
S2: Additional NMR characterization of J2LS fragments.Predicted secondary structure of JR2 VBPs as calculated based on the
backbone and side chain NMR chemical shifts using the TALOS algorithm.
S3: Additional NMR data . A) Data table showing various
parameters relating to the NMR structure determination and validation of
A1pro and JR2 LS fragments. B) Cysteine CB chemical shift values for all
four constructs. Values indicate that all cystines are involved in
disulfide bonds.
S4: Surface comparison with distant VBP’s. A) NMR structure of
VBP-8; and B) BWI-2C VBP’s from tomato and buckwheat respectively. Spade
surface-similarity comparison with immunodominant A1LS and JR2.1. shown
below, revealing areas of high surface similarity despite the lack of
known tomato-nut and buckwheat-nut cross-reactivity . PDB ID: 6O3S, 2LQX