Figure 1.
Bombykol receptor expressed in the Xenopus oocyte recording system.
Robust currents from BmorOR1•BmorOrco-expressing oocytes when perfused with bombykol, and dose-dependent responses. n = 3–5, error bars in all figures represent SEM.
Figure 2.
(A) Current responses and (B) dose-dependent relationships obtained by challenging BmorOR1•BmorOrco-expressing oocytes with increasing concentrations of bombykol and bombykal. n = 6.
Figure 3.
Chemical analysis of synthetic pheromone components.
GC-MS traces obtained from bombykol (upper trace) and bombykal (lower trace) samples freshly prepared to challenge BmorOR1•BmorOrco-expressing oocytes. Arrow indicates trace amounts of bombykal (1.3%) in the bombykol sample, whereas a dotted arrow shows traces of bombykol (0.9%) in bombykal sample. The ratio of bombykol (retention time, 16.06 min) to bombykal (15.42 min) in the two samples was 1.015±0.02, n = 3.
Figure 4.
Synthetic pheromone components trapped by PBP.
(A) Traces and (B) quantification of current responses obtained from the BmorOR1•BmorOrco-expressing oocytes when presented with bombykol and bombykal solubilized either by DMSO or BmorPBP1. n = 3. *Significantly different (t-test, P<0.05).
Figure 5.
Structures of the silkworm moth sex pheromone (1) and bombykol-related compounds, which were used to challenge BmorOR1•BmorOrco-expressing oocytes.
Figure 6.
(A) Traces and (B) quantification of current responses from BmorOR1•BmorOrco-expressing oocytes perfused with four isomers of bombykol at 0.1 µM. n = 5. Bars with the same letter arwe not significantly different (One-way ANOVA, P<0.01).
Figure 7.
Effect of altering unsaturation on receptor response.
(A) Traces and (B) quantification of current responses elicited by (8E,10Z)-hexadecadien-1-ol (9) and 10,12-hexadecadiyn-1-ol (6) presented at 1 mM. n = 3. Bars with the same letter are not significantly different (One-way ANOVA, P<0.01).
Figure 8.
Effect of number of carbons distal to unsaturation.
(A) Current responses obtained by challenging BmorOR1•BmorOrco-expressing oocytes with bombykol (lower trace, positive control), (10E,12Z)-octadecadien-1-ol (7), and (10E,12Z)-tetradecadien-1-ol (8), robust response at 10 µM. (B) Dose-dependent relationships; n = 4.
Figure 9.
Reducing responses by adding rigidity to the C1–C9 moiety.
(A) Current responses elicited by (10E,12Z)-hexadecadien-4-yn-1-ol (10), (4Z,10E,12Z)-hexadecatrien-1-ol (11), and bombykol (1) from BmorOR1•BmorOrco-expressing oocytes (ligands presented at 10 µM). EC50s 1.7×10−5M, 1.3×10−5M, and 5.9×10−6M, respectively. (B) Dose-dependent relationships, n = 3–4.
Figure 10.
Synthetic sequence for preparation of analogues 7–11 containing the (E,Z)-dienyl moiety.
Figure 11.
Synthetic sequence for preparation of analogues 3–6 differing in unsaturation.