Evolving the N-Terminal Domain of Pyrrolysyl-tRNA Synthetase for Improved Incorporation of Noncanonical Amino Acids

Chembiochem : a European Journal of Chemical Biology
Vangmayee SharmaWenshe R Liu

Abstract

By evolving the N-terminal domain of Methanosarcina mazei pyrrolysyl-tRNA synthetase (PylRS) that directly interacts with tRNAPyl , a mutant clone displaying improved amber-suppression efficiency for the genetic incorporation of Nϵ -(tert-butoxycarbonyl)-l-lysine threefold more than the wild type was identified. The identified mutations were R19H/H29R/T122S. Direct transfer of these mutations to two other PylRS mutants that were previously evolved for the genetic incorporation of Nϵ -acetyl-l-lysine and Nϵ -(4-azidobenzoxycarbonyl)-l-δ,ϵ-dehydrolysine also improved the incorporation efficiency of these two noncanonical amino acids. As the three identified mutations were found in the N-terminal domain of PylRS that was separated from its catalytic domain for charging tRNAPyl with a noncanonical amino acid, they could potentially be introduced to all other PylRS mutants to improve the incorporation efficiency of their corresponding noncanonical amino acids. Therefore, it represents a general strategy to optimize the pyrrolysine incorporation system-based noncanonical amino-acid mutagenesis.

References

Jun 18, 1999·Methods in Enzymology·K LugerT J Richmond
May 25, 2002·Science·Gayathri SrinivasanJoseph A Krzycki
Aug 18, 2004·Proceedings of the National Academy of Sciences of the United States of America·Carla PolycarpoDieter Söll
Aug 27, 2004·Nature·Sherry K BlightJoseph A Krzycki
Nov 4, 2004·Journal of the American Chemical Society·Ning WuPeter G Schultz
Nov 28, 2006·FEBS Letters·Carla R PolycarpoAlexandre Ambrogelly
Feb 3, 2007·Nucleic Acids Research·Stephanie HerringDieter Söll
Mar 16, 2007·Proceedings of the National Academy of Sciences of the United States of America·Alexandre AmbrogellyDieter Söll
Jun 27, 2007·Proceedings of the National Academy of Sciences of the United States of America·Jennifer M KavranThomas A Steitz
Oct 31, 2007·FEBS Letters·Olivier NamyAlexandre Ambrogelly
Feb 19, 2008·Nature Chemical Biology·Heinz NeumannJason W Chin
May 13, 2008·Biochemical and Biophysical Research Communications·Takahito MukaiShigeyuki Yokoyama
Oct 27, 2009·Journal of Molecular Biology·Travis S YoungPeter G Schultz
Mar 12, 2010·Journal of the American Chemical Society·Arnaud GautierJason W Chin
Aug 9, 2011·Journal of the American Chemical Society·Sebastian Greiss, Jason W Chin
Sep 6, 2011·Nature Chemical Biology·Meng ZhangPeng R Chen
Apr 5, 2012·Angewandte Chemie·Tilman PlassEdward A Lemke
Mar 19, 2014·Biochimica Et Biophysica Acta·Wei WanWenshe R Liu
Aug 8, 2015·Nucleic Acids Research·Chenguang FanDieter Söll
Jan 29, 2016·ACS Chemical Biology·Willie W HsuWenshe R Liu
Mar 16, 2016·Proceedings of the National Academy of Sciences of the United States of America·Xiaozhou LuoTravis S Young
May 7, 2016·ACS Chemical Biology·Wesley Wei WangWenshe R Liu
Apr 7, 2017·Chembiochem : a European Journal of Chemical Biology·Andrew E OwensRudi Fasan

❮ Previous
Next ❯

Citations

Oct 31, 2019·Chembiochem : a European Journal of Chemical Biology·Joshua S WesaloAlexander Deiters
Oct 1, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Hsueh-Wei TsengNediljko Budisa
May 15, 2019·International Journal of Molecular Sciences·Tobias BaumannNediljko Budisa
Apr 24, 2020·Frontiers in Bioengineering and Biotechnology·Han-Kai JiangYane-Shih Wang
Mar 24, 2021·Journal of the American Chemical Society·Miglena ManandharFloyd E Romesberg

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.