Virus-Enabled Optimization and Delivery of the Genetic Machinery for Efficient Unnatural Amino Acid Mutagenesis in Mammalian Cells and Tissues

2016 ◽  
Vol 6 (1) ◽  
pp. 13-18 ◽  
Author(s):  
Yunan Zheng ◽  
Tommy L. Lewis ◽  
Peter Igo ◽  
Franck Polleux ◽  
Abhishek Chatterjee
Keyword(s):  
Amino Acid ◽  
Mammalian Cells ◽  
Unnatural Amino Acid ◽  
Unnatural Amino Acid Mutagenesis ◽  
Amino Acid Mutagenesis

scholarly journals Gating modules of the AMPA receptor pore domain revealed by unnatural amino acid mutagenesis

2019 ◽  
Vol 116 (27) ◽  
pp. 13358-13367 ◽  
Author(s):  
Mette H. Poulsen ◽  
Anahita Poshtiban ◽  
Viktoria Klippenstein ◽  
Valentina Ghisi ◽  
Andrew J. R. Plested
Keyword(s):  
Amino Acid ◽  
Conformational Changes ◽  
Transmembrane Domain ◽  
Uv Light ◽  
Unnatural Amino Acid ◽  
Selectivity Filter ◽  
Unnatural Amino Acid Mutagenesis ◽  
Amino Acid Mutagenesis ◽  
Mutant Receptors ◽  
Pore Domain

Ionotropic glutamate receptors (iGluRs) are responsible for fast synaptic transmission throughout the vertebrate nervous system. Conformational changes of the transmembrane domain (TMD) underlying ion channel activation and desensitization remain poorly understood. Here, we explored the dynamics of the TMD of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type iGluRs using genetically encoded unnatural amino acid (UAA) photocross-linkers, p-benzoyl-l-phenylalanine (BzF) and p-azido-l-phenylalanine (AzF). We introduced these UAAs at sites throughout the TMD of the GluA2 receptor and characterized the mutants in patch-clamp recordings, exposing them to glutamate and ultraviolet (UV) light. This approach revealed a range of optical effects on the activity of mutant receptors. We found evidence for an interaction between the Pre-M1 and the M4 TMD helix during desensitization. Photoactivation at F579AzF, a residue behind the selectivity filter in the M2 segment, had extraordinarily broad effects on gating and desensitization. This observation suggests coupling to other parts of the receptor and like in other tetrameric ion channels, selectivity filter gating.

scholarly journals Investigation of Trimethyllysine Binding by the HP1 Chromodomain via Unnatural Amino Acid Mutagenesis

2017 ◽  
Vol 139 (48) ◽  
pp. 17253-17256 ◽  
Author(s):  
Stefanie A. Baril ◽  
Amber L. Koenig ◽  
Mackenzie W. Krone ◽  
Katherine I. Albanese ◽  
Cyndi Qixin He ◽  
...  
Keyword(s):  
Amino Acid ◽  
Unnatural Amino Acid ◽  
Unnatural Amino Acid Mutagenesis ◽  
Amino Acid Mutagenesis
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