INVESTIGADORES
BOUZAT Cecilia Beatriz
congresos y reuniones científicas
Título:
Contribution of agonist binding sites and coupling regions to activation and desensitization of homomeric Cys-loop receptors. Symposium: Ligand-Gated Ion Channels
Autor/es:
BOUZAT, C.
Reunión:
Conferencia; 56th Annual Meeting of the Biophysical Society; 2012
Resumen:
Homomeric receptors are the simplest structural class of receptors of the Cys-loop superfamily andare therefore invaluable models for probing fundamental relationships between structure andfunction. To determine how many of the five agonist binding sites are required to be occupied bythe agonist for maximal open channel stability we applied an electrical fingerprinting strategy in thehomomeric receptor model, α7-5HT3A. To vary the number of functional agonist binding sites weinstalled mutations that prevent agonist binding, and to report the presence of the mutant subunit,we installed mutations that alter single-channel conductance. We find that receptors can beactivated by occupancy of only one agonist binding site but open channel lifetime is brief, andoccupancy of three non-consecutive sites is required for maximal open channel lifetime. Theconformational changes initiated at the binding site are propagated to the gate through theextracellular-transmembrane interface, known as coupling region. We show that structuraldifferences in the coupling region of homomeric α7 and 5-HT3A receptors account for the largedifferences in open channel lifetime and rate of desensitization between these homomeric members of the superfamily. By applying the electrical fingerprinting strategy, we determine that each coupling region in the pentamer contributes an equal increment to the stability of the open channel. We also determine minimal requirements for channel opening regardless of stability of the open state, and find that receptors can open with one functional binding site and two contiguous and functional coupling regions, or with five functional binding sites and only one functional coupling region. The overall findings show that whereas the agonist binding sites contribute interdependently and asymmetrically to open channel stability, the coupling regions contributeindependently and symmetrically.