scholarly journals Retroviral gag gene amber codon suppression is caused by an intrinsic cis-acting component of the viral mRNA.

1988 ◽  
Vol 62 (10) ◽  
pp. 3574-3580 ◽  
Author(s):  
A T Panganiban
Keyword(s):  
Viral Mrna ◽  
Cis Acting ◽  
Amber Codon ◽  
Amber Codon Suppression

scholarly journals Site-Specific Labelling of Multidomain Proteins by Amber Codon Suppression

2018 ◽  
Author(s):  
Christina S. Heil ◽  
Alexander Rittner ◽  
Bjarne Goebel ◽  
Daniel Beyer ◽  
Martin Grininger
Keyword(s):  
Fatty Acid ◽  
Fatty Acid Synthase ◽  
Acyl Carrier Protein ◽  
Protein Domain ◽  
Type I ◽  
Reporter Assay ◽  
Multidomain Proteins ◽  
Conformational Variability ◽  
Amber Codon ◽  
Amber Codon Suppression

AbstractAmber codon suppression is a powerful tool to site-specifically modify proteins to generate novel biophysical probes. Yet, its application on large and complex multidomain proteins is challenging, leading to difficulties during structural and conformational characterization using spectroscopic methods. The animal fatty acid synthase type I is a 540 kDa homodimer displaying large conformational variability. As the key enzyme of de novo fatty acid synthesis, it attracts interest in the fields of obesity, diabetes and cancer treatment. Substrates and intermediates remain covalently bound to the enzyme during biosynthesis and are shuttled to all catalytic domains by the acyl carrier protein domain. Thus, conformational variability of animal FAS is an essential aspect for fatty acid biosynthesis. We investigate this multidomain protein as a model system for probing amber codon suppression by genetic encoding of non-canonical amino acids. The systematic approach relies on a microplate-based reporter assay of low complexity, that was used for quick screening of suppression conditions. Furthermore, the applicability of the reporter assay is demonstrated by successful upscaling to both full-length constructs and increased expression scale. The obtained fluorescent probes of murine FAS type I could be subjected readily to a conformational analysis using single-molecule fluorescence resonance energy transfer.

Tag-on-Demand: exploiting amber codon suppression technology for the enrichment of high-expressing membrane protein cell lines

2018 ◽  
Vol 31 (10) ◽  
pp. 389-398 ◽  
Author(s):  
Zachary T Britton ◽  
Timothy B London ◽  
Jeffrey Carrell ◽  
Bhupinder Dosanjh ◽  
Trevor Wilkinson ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Cell Lines ◽  
On Demand ◽  
Amber Codon ◽  
Amber Codon Suppression ◽  
Protein Cell

Synthesis of post-translationally modified proteins

2012 ◽  
Vol 40 (5) ◽  
pp. 929-944 ◽  
Author(s):  
Sander van Kasteren
Keyword(s):  
Research Effort ◽  
Native Chemical Ligation ◽  
Chemical Ligation ◽  
Post Translational Modifications ◽  
Dramatic Effect ◽  
Amber Codon ◽  
Significant Research ◽  
Modified Proteins ◽  
Amber Codon Suppression

Post-translational modifications of proteins can have dramatic effect on the function of proteins. Significant research effort has gone into understanding the effect of particular modifications on protein parameters. In the present paper, I review some of the recently developed tools for the synthesis of proteins modified with single post-translational modifications at specific sites in the protein, such as amber codon suppression technologies, tag and modify, and native chemical ligation.

Amber codon suppression: the in vivo and in vitro analysis of two C-hordein genes from barley

1991 ◽  
Vol 17 (6) ◽  
pp. 1217-1231 ◽  
Author(s):  
Joycelyn Entwistle ◽  
S�ren Knudsen ◽  
Martin M�ller ◽  
Verena Cameron-Mills
Keyword(s):  
Amber Codon ◽  
Vitro Analysis ◽  
Amber Codon Suppression ◽  
In Vitro Analysis

scholarly journals Site-Specific Labelling of Multidomain Proteins by Amber Codon Suppression

2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Christina S. Heil ◽  
Alexander Rittner ◽  
Bjarne Goebel ◽  
Daniel Beyer ◽  
Martin Grininger
Keyword(s):  
Multidomain Proteins ◽  
Site Specific ◽  
Amber Codon ◽  
Amber Codon Suppression ◽  
Specific Labelling

scholarly journals Genetic incorporation of 1,2-aminothiol functionality for site-specific protein modification via thiazolidine formation

2016 ◽  
Vol 14 (23) ◽  
pp. 5282-5285 ◽  
Author(s):  
Xiaobao Bi ◽  
Kalyan Kumar Pasunooti ◽  
Ahmad Hussen Tareq ◽  
John Takyi-Williams ◽  
Chuan-Fa Liu
Keyword(s):  
Protein Modification ◽  
Specific Protein ◽  
Site Specific ◽  
Amber Codon ◽  
Amber Codon Suppression ◽  
Genetic Incorporation

Thiazolidine ligation was used to modify site-specifically proteins harbouring a 1,2-aminothiol moiety introduced by amber codon suppression technology.

scholarly journals Photosensitive tyrosine analogues unravel site-dependent phosphorylation in TrkA initiated MAPK/ERK signaling

2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Shu Zhao ◽  
Jia Shi ◽  
Guohua Yu ◽  
Dali Li ◽  
Meng Wang ◽  
...  
Keyword(s):  
Ligand Binding ◽  
Phosphorylation Site ◽  
Optical Control ◽  
Erk Signaling ◽  
Kinase Activation ◽  
Amber Codon ◽  
Genetic Code Expansion ◽  
A Site ◽  
Amber Codon Suppression ◽  
Signaling Process

AbstractTyrosine kinase A (TrkA) is a membrane receptor which, upon ligand binding, activates several pathways including MAPK/ERK signaling, implicated in a spectrum of human pathologies; thus, TrkA is an emerging therapeutic target in treatment of neuronal diseases and cancer. However, mechanistic insights into TrKA signaling are lacking due to lack of site-dependent phosphorylation control. Here we engineer two light-sensitive tyrosine analogues, namely p-azido-L-phenylalanine (AzF) and the caged-tyrosine (ONB), through amber codon suppression to optically manipulate the phosphorylation state of individual intracellular tyrosines in TrkA. We identify TrkA-AzF and ONB mutants, which can activate the ERK pathway in the absence of NGF ligand binding through light control. Our results not only reveal how TrkA site-dependent phosphorylation controls the defined signaling process, but also extend the genetic code expansion technology to enable regulation of receptor-type kinase activation by optical control at the precision of a single phosphorylation site. It paves the way for comprehensive analysis of kinase-associated pathways as well as screening of compounds intervening in a site-directed phosphorylation pathway for targeted therapy.

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