cn500369h_si_002.pdf (2.92 MB)
Varenicline Interactions at the 5‑HT3 Receptor Ligand Binding Site are Revealed by 5‑HTBP
journal contribution
posted on 2015-12-17, 08:41 authored by Kerry
L. Price, Reidun K. Lillestol, Chris Ulens, Sarah C.R. LummisCys-loop receptors are the site of
action of many therapeutic drugs.
One of these is the smoking cessation agent varenicline, which has
its major therapeutic effects at nicotinic acetylcholine (nACh) receptors
but also acts at 5-HT3 receptors. Here, we report the X-ray
crystal structure of the 5-HT binding protein (5-HTBP) in complex
with varenicline, and test the predicted interactions by probing the
potency of varenicline in a range of mutant 5-HT3 receptors
expressed in HEK293 cells and Xenopus oocytes. The
structure reveals a range of interactions between varenicline and
5-HTBP. We identified residues within 5 Å of varenicline and
substituted the equivalent residues in the 5-HT3 receptor
with Ala or a residue with similar chemical properties. Functional
characterization of these mutant 5-HT3 receptors, using
a fluorescent membrane potential dye in HEK cells and voltage clamp
in oocytes, supports interactions between varenicline and the receptor
that are similar to those in 5-HTBP. The structure also revealed C-loop
closure that was less than in the 5-HT-bound 5-HTBP, and hydrogen
bonding between varenicline and the complementary face of the binding
pocket via a water molecule, which are characteristics consistent
with partial agonist behavior of varenicline in the 5-HT3 receptor. Together, these data reveal detailed insights into the
molecular interaction of varenicline in the 5-HT3 receptor.