Supplementary Figure 5: Characterization of binding between HCA variants and human bSV2C and gSV2C.
From: N-linked glycosylation of SV2 is required for binding and uptake of botulinum neurotoxin A
(a) Surface plasmon resonance was used to examine the changes of binding affinity between HCA variants and SUMO-bSV2C or gSV2C, respectively. SV2C was covalently immobilized to a CM5 chip as a ligand whereas HCA variants were analytes. Bars from left to right represent the responses when HCA was applied at 10 pM, 1 nM, 10 nM, 25 nM, 50 nM, 75 nM, 100 nM, and 200 nM, respectively. RU stands for arbitrary response unit. (b,c) Interactions between HCA variants (preys) and SUMO-bSV2C or gSV2C (baits) were examined by a pull down assay. (d,e) Binding kinetics and affinity between HCA-F953G and immobilized bSV2C (107 RU; panel d) or gSV2C (74 RU; panel e) were determined by injecting 1:3 dilution series ranging from 2,000 nM to 8.23 nM. Values shown represent the mean ±S.D. (n = 2).
