Section 55
Chapter 54,985

Photoinactivation of glutamate receptors by genetically encoded unnatural amino acids

Klippenstein, V.; Ghisi, V.; Wietstruk, M.; Plested, A.J.R.

Journal of Neuroscience the Official Journal of the Society for Neuroscience 34(3): 980-991


ISSN/ISBN: 1529-2401
PMID: 24431456
DOI: 10.1523/jneurosci.3725-13.2014
Accession: 054984650

Download citation:  

Ionotropic glutamate receptors (iGluRs) are ubiquitous in the mammalian brain, and the AMPA-subtype is essential for fast, glutamate-activated postsynaptic currents. We incorporated photoactive crosslinkers into AMPA receptors using genetically encoded unnatural amino acid mutagenesis in a mammalian cell line. Receptors rescued by incorporation of unnatural amino acids, including p-benzoyl-l-phenylalanine (BzF, also known as Bpa), had largely similar properties to wild-type channels and were expressed at similar levels. BzF incorporation at subunit interfaces afforded photocrosslinking of subunits, as assessed by biochemical experiments. In electrophysiological recordings, BzF incorporation allowed selective and potent UV-driven photoinactivation of both homomeric (GluA2) and heteromeric (GluA2:GluA1) AMPA receptors. State dependence of trapping at two sites in the lower lobe of the ligand binding domain is consistent with deformation of these domains as well as intersubunit rearrangements during AMPA receptor desensitization.

PDF emailed within 0-6 h: $19.90