How AMPA receptor desensitization depends on receptor occupancy.
AMPA-type glutamate receptors mediate fast excitatory transmission at many central synapses, and rapid desensitization of these receptors can shape the decay of synaptic currents and limit the fidelity of high-frequency synaptic transmission. Here we use a combination of fast glutamate application protocols and kinetic simulations to determine how AMPA receptor desensitization depends on the number of subunits occupied by glutamate. We show that occupancy of a single subunit is sufficient to desensitize AMPA-type channels and that receptors with one to four glutamates bound enter desensitization at similar rates. We find that recovery from desensitization follows a similar sigmoid time course for channels with two to four glutamates bound but is faster and exponential for singly occupied channels. The results suggest that desensitization, at intermediate and high glutamate concentrations, is accompanied by two conformational changes that slow glutamate dissociation. We propose a kinetic scheme that accurately predicts several types of experimental results and differs significantly from previous models in the assignment of affinities for binding to closed and desensitized states. We conclude that desensitization involves a rearrangement that stabilizes the binding domains of one subunit in each dimer in a partially closed conformation. This stabilization likely results from an interaction at the dimer-dimer interface between the binding domains of adjacent subunits.
Robert
Antoine
A
Department of Pharmacology, Yale University School of Medicine, New Haven, Connecticut 06520-8066, USA.
Howe
James R
JR
D001665:Binding Sites / Q000502:physiology; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D018846:Excitatory Amino Acids / Q000493:pharmacokinetics / Q000494:pharmacology*; D018698:Glutamic Acid / Q000493:pharmacokinetics / Q000494:pharmacology*; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D007700:Kinetics; D008962:Models, Theoretical; D018408:Patch-Clamp Techniques; D010477:Perfusion / Q000379:methods; D011487:Protein Conformation / Q000187:drug effects; D018091:Receptors, AMPA / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D018846:Excitatory Amino Acids; D018698:Glutamic Acid; D018091:Receptors, AMPA; C104724:glutamate receptor ionotropic, AMPA 4