| Title | Optocontrol of glutamate receptor activity by single side-chain photoisomerization |
| Author | Viktoria Klippenstein1, Christian Hoppmann2,3, Shixin Ye1,4, Lei Wang2,3, Pierre Paoletti1* |
| Affiliation(s) | 1 Institut de Biologie de l’E ´cole Normale Supe´rieure, Ecole Normale Supe´rieure, CNRS, INSERM, PSL Research University, Paris, France; 2 Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, United States; 3 Cardiovascular Research Institute, University of California, San Francisco, San Francisco, United States; 4 Laboratory of Computational and Quantitative Biology, Universite´ Pierre-et-Marie-Curie, CNRS, Paris, France |
| Published | eLife, 2017 - elifesciences.org DOI: 10.7554/eLife.25808.001 |
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| Abstract | Engineering light-sensitivity into proteins has wide ranging applications in molecular studies and neuroscience. Commonly used tethered photoswitchable ligands, however, require solvent-accessible protein labeling, face structural constrains, and are bulky. Here, we designed a set of optocontrollable NMDA receptors by directly incorporating single photoswitchable amino acids (PSAAs) providing genetic encodability, reversibility, and site tolerance. We identified several positions within the multi-domain receptor endowing robust photomodulation. PSAA photoisomerization at the GluN1 clamshell hinge is sufficient to control glycine sensitivity and activation efficacy. Strikingly, in the pore domain, flipping of a M3 residue within a conserved transmembrane cavity impacts both gating and permeation properties. Our study demonstrates the first detection of molecular rearrangements in real-time due to the reversible light-switching of single amino acid side-chains, adding a dynamic dimension to protein site-directed mutagenesis. This novel approach to interrogate neuronal protein function has general applicability in the fast expanding field of optopharmacology |
