Dynamics of cleft closure of the GluA2 ligand-binding domain in the presence of full and partial agonists revealed by hydrogen-deuterium exchange.
The majority of excitatory neurotransmission in the CNS is mediated by tetrameric AMPA receptors. Channel activation begins with a series of interactions with an agonist that binds to the cleft between the two lobes of the ligand-binding domain of each subunit. Binding leads to a series of conformational transitions, including the closure of the two lobes of the binding domain around the ligand, culminating in ion channel opening. Although a great deal has been learned from crystal structures, determining the molecular details of channel activation, deactivation, and desensitization requires measures of dynamics and stabilities of hydrogen bonds that stabilize cleft closure. The use of hydrogen-deuterium exchange at low pH provides a measure of the variation of stability of specific hydrogen bonds among agonists of different efficacy. Here, we used NMR measurements of hydrogen-deuterium exchange to determine the stability of hydrogen bonds in the GluA2 (AMPA receptor) ligand-binding domain in the presence of several full and partial agonists. The results suggest that the stabilization of hydrogen bonds between the two lobes of the binding domain is weaker for partial than for full agonists, and efficacy is correlated with the stability of these hydrogen bonds. The closure of the lobes around the agonists leads to a destabilization of the hydrogen bonding in another portion of the lobe interface, and removing an electrostatic interaction in Lobe 2 can relieve the strain. These results provide new details of transitions in the binding domain that are associated with channel activation and desensitization.
Departments of Molecular Medicine.
Ahmed H
AH
Ahmed
Departments of Molecular Medicine.
Christopher P
CP
Ptak
Chemistry and Chemical Biology.
Michael K
MK
Fenwick
Population Medicine and Diagnostic Sciences, Cornell University, Ithaca, New York 14853.
Ching-Lin
CL
Hsieh
Departments of Molecular Medicine.
Gregory A
GA
Weiland
Departments of Molecular Medicine. Electronic address: reo1@cornell.edu.
Robert E
RE
Oswald
D000818:Animals; D041961:Deuterium Exchange Measurement / Q000379:methods; D006860:Hydrogen Bonding; D006863:Hydrogen-Ion Concentration; D056004:Molecular Dynamics Simulation; D017434:Protein Structure, Tertiary; D051381:Rats; D018091:Receptors, AMPA / Q000819:agonists* / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship
D018091:Receptors, AMPA; C104722:glutamate receptor ionotropic, AMPA 2
Glutamate Receptors; Hydrogen-Deuterium Exchange; Ionotropic Glutamate Receptors (AMPA, NMDA); Membrane Proteins; NMR; Neurotransmitter Receptors; Willardiine