In the Plested lab, we study the molecular basis of neurotransmitter receptor activation. Our main interest is the glutamate receptors that mediate excitatory transmission at ~ 60% of synapses in the brain, but we are also interested in other ligand gated ion channels. We study these channels because they are essential for nervous system function, and because they are fascinating and complex protein machines.
We investigate the function of glutamate receptors using patch clamp electrophysiology, single channel recording and ultrarapid perfusion techniques. We employ chemical modification of cloned channels, and map receptor activation with disulfide crosslinks and metal bridges. We also overexpress and purify receptor domains for biophysical experiments and crystallization, to investigate the structural bases of receptor activation. To achieve this aim, we pursue collaborations with groups at MDC, and work at the local BESSY synchrotron. Through these techniques we aim to infer information about the dynamics and assembly of the glutamate receptor.
A second line of attack involves extending these approaches to other components of fast signaling in the brain including voltage -gated ion channels and inhibitory neurotransmitter receptors.
Lau AY†*, Salazar H*, Blachowitz L, Ghisi V, Plested AJR†, Roux B† (2013)
A conformational intermediate in glutamate receptor activation. Neuron
*These authors contributed jointly. †Corresponding authors
Miranda P, Contreras JE, Plested AJR, Sigworth FJ, Holmgren M, and Giraldez T (2013)
State-dependent FRET reports calcium and voltage-dependent gating-ring motions in whole BK channels
PNAS 110(13): 5217-22
Carbone AL and Plested AJR (2012)
Coupled control of desensitization and gating by the ligand binding domain of glutamate receptors
Neuron 74: 845–857
Lape R*, Plested AJR*, Moroni M, Colquhoun D and Sivilotti LG (2012)
The α1K276E startle disease mutation reveals multiple intermediate states in the gating of glycine receptors.
Journal of Neuroscience 32(4):1336-52.
*These authors contributed jointly.
Das U, Kumar J, Mayer ML† and Plested AJR† (2010)
Domain organization and function in GluK2 subtype kainate receptors.