Fig. 2: Interaction and functional effects of the HSV1 gD whole ectodomain with L-AChBP and the α7 nAChR.
From: HSV1 glycoprotein D utilizes an LY6-like binding domain to inhibit alpha7 nicotinic receptors
a HSV1 gD whole ectodomain binding to the L -AChBP. SPR was conducted using four different concentrations of HSV1 gD whole ectodomain (7.8 nM, 31 nM, 1.2 μM and 5 μM). L-AChBP protein was immobilized on the CM chip surface (the ligand) and HSV1 gD was flowed over the chip surface (the analyte). Analyte was injected for 4 min at a flow rate of 25 μL/min. Affinity was calculated using a 1:1 binding model (Kd = 2.12 × 10-6 ± 5.31 × 10-7 M). b Functional effects of HSV1 gD on α7 nAChRs expressed in Xenopus oocytes. Oocytes were injected with mRNA coding for the human α7 nAChR. Oocytes were then co-exposed to both nicotine (30 μM) and the 319 AA whole ectodomain of HSV1 gD. 30 μM nicotine produced approximately a 40% Imax response that was inhibited by increasing concentrations of HSV1 gD. The four different concentrations of HSV1 gD shown (0.1 μM, 0.33 μM, 1 μM, 2 μM) produced an area under the curve (total current) of 502, 331, 138, and 10 uA·ms respectively. c Dose response curve for inhibition of nicotinic induced responses by HSV1 gD. The AUC data obtained from oocyte inhibition experiments was used to create an inhibitory dose response curve for HSV1 gD. Data from at least 4 oocytes was pooled to create the curve shown. IC50 = 0.85 µM (95% CI: 0.55 to 1.14 µM).
