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

Unnatural amino acid mutagenesis in mapping ion channel function

2003 ◽  
Vol 13 (3) ◽  
pp. 264-270 ◽  
Author(s):  
Darren L Beene ◽  
Dennis A Dougherty ◽  
Henry A Lester
Keyword(s):  
Amino Acid ◽  
Ion Channel ◽  
Unnatural Amino Acid ◽  
Channel Function ◽  
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.

Analysis of the Role of the Active Site Tyrosine in Human Glutathione Transferase A1-1 by Unnatural Amino Acid Mutagenesis

1998 ◽  
Vol 120 (2) ◽  
pp. 451-452 ◽  
Author(s):  
Jon S. Thorson ◽  
Injae Shin ◽  
Eli Chapman ◽  
Gun Stenberg ◽  
Bengt Mannervik ◽  
...  
Keyword(s):  
Amino Acid ◽  
Active Site ◽  
Glutathione Transferase ◽  
Unnatural Amino Acid ◽  
Unnatural Amino Acid Mutagenesis ◽  
Active Site Tyrosine ◽  
Amino Acid Mutagenesis

scholarly journals A Novel Method to Probe Membrane Protein Topology Using Unnatural Amino Acid Mutagenesis and Antibody Epitope Tagging

2010 ◽  
Vol 98 (3) ◽  
pp. 419a
Author(s):  
Saranga Naganathan ◽  
Shixin Ye ◽  
Terence Duarte ◽  
Thomas Huber ◽  
Pallavi Sachdev ◽  
...  
Keyword(s):  
Amino Acid ◽  
Membrane Protein ◽  
Unnatural Amino Acid ◽  
Epitope Tagging ◽  
Protein Topology ◽  
Unnatural Amino Acid Mutagenesis ◽  
Membrane Protein Topology ◽  
Antibody Epitope ◽  
Novel Method ◽  
Amino Acid Mutagenesis

scholarly journals Testing Models for the Slow Inactivated State using Unnatural Amino Acid Mutagenesis

2012 ◽  
Vol 102 (3) ◽  
pp. 530a
Author(s):  
Francis Valiyaveetil ◽  
Alexander Komarov ◽  
Prasanna Devaraneni
Keyword(s):  
Amino Acid ◽  
Unnatural Amino Acid ◽  
Unnatural Amino Acid Mutagenesis ◽  
Amino Acid Mutagenesis

scholarly journals Site-specific Incorporation of Keto Amino Acids into Functional G Protein-coupled Receptors Using Unnatural Amino Acid Mutagenesis

2007 ◽  
Vol 283 (3) ◽  
pp. 1525-1533 ◽  
Author(s):  
Shixin Ye ◽  
Caroline Köhrer ◽  
Thomas Huber ◽  
Manija Kazmi ◽  
Pallavi Sachdev ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
G Protein ◽  
Unnatural Amino Acids ◽  
Unnatural Amino Acid ◽  
G Protein Coupled Receptors ◽  
Calcium Flux ◽  
Unnatural Amino Acid Mutagenesis ◽  
Amino Acid Mutagenesis ◽  
G Protein Coupled

G protein-coupled receptors (GPCRs) are ubiquitous heptahelical transmembrane proteins involved in a wide variety of signaling pathways. The work described here on application of unnatural amino acid mutagenesis to two GPCRs, the chemokine receptor CCR5 (a major co-receptor for the human immunodeficiency virus) and rhodopsin (the visual photoreceptor), adds a new dimension to studies of GPCRs. We incorporated the unnatural amino acids p-acetyl-l-phenylalanine (Acp) and p-benzoyl-l-phenylalanine (Bzp) into CCR5 at high efficiency in mammalian cells to produce functional receptors harboring reactive keto groups at three specific positions. We obtained functional mutant CCR5, at levels up to ∼50% of wild type as judged by immunoblotting, cell surface expression, and ligand-dependent calcium flux. Rhodopsin containing Acp at three different sites was also purified in high yield (0.5–2 μg/107 cells) and reacted with fluorescein hydrazide in vitro to produce fluorescently labeled rhodopsin. The incorporation of reactive keto groups such as Acp or Bzp into GPCRs allows their reaction with different reagents to introduce a variety of spectroscopic and other probes. Bzp also provides the possibility of photo-cross-linking to identify precise sites of protein-protein interactions, including GPCR binding to G proteins and arrestins, and for understanding the molecular basis of ligand recognition by chemokine receptors.

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