Design ofS-Allylcysteine in Situ Production and Incorporation Based on a Novel Pyrrolysyl-tRNA Synthetase Variant
| Autor: | Sergej Schwagerus, Marga C. Lensen, Zhaofei Ouyang, Tilmann Kuenzl, Nediljko Budisa, Sven Panke, Michael Georg Hoesl, Tuyet Mai T. To, Christian P. R. Hackenberger, Matthias P. Exner |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
Stereochemistry
Green Fluorescent Proteins Protein design Biology 010402 general chemistry 01 natural sciences Biochemistry Amino Acyl-tRNA Synthetases chemistry.chemical_compound Biosynthesis Escherichia coli Cysteine Molecular Biology chemistry.chemical_classification Cysteine Synthase Binding Sites 010405 organic chemistry Organic Chemistry Translation (biology) Stop codon Protein Structure Tertiary 0104 chemical sciences Amino acid Molecular Docking Simulation chemistry Methanosarcina Transfer RNA Mutagenesis Site-Directed Molecular Medicine Bioorthogonal chemistry |
| Zdroj: | ChemBioChem. 18:85-90 |
| ISSN: | 1439-4227 |
| DOI: | 10.1002/cbic.201600537 |
| Popis: | The noncanonical amino acid S-allyl cysteine (Sac) is one of the major compounds of garlic extract and exhibits a range of biological activities. It is also a small bioorthogonal alkene tag capable of undergoing controlled chemical modifications, such as photoinduced thiol-ene coupling or Pd-mediated deprotection. Its small size guarantees minimal interference with protein structure and function. Here, we report a simple protocol efficiently to couple in-situ semisynthetic biosynthesis of Sac and its incorporation into proteins in response to amber (UAG) stop codons. We exploited the exceptional malleability of pyrrolysyl-tRNA synthetase (PylRS) and evolved an S-allylcysteinyl-tRNA synthetase (SacRS) capable of specifically accepting the small, polar amino acid instead of its long and bulky aliphatic natural substrate. We succeeded in generating a novel and inexpensive strategy for the incorporation of a functionally versatile amino acid. This will help in the conversion of orthogonal translation from a standard technique in academic research to industrial biotechnology. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text