| RRC ID |
69153
|
| 著者 |
Thyer R, d'Oelsnitz S, Blevins MS, Klein DR, Brodbelt JS, Ellington AD.
|
| タイトル |
Directed Evolution of an Improved Aminoacyl-tRNA Synthetase for Incorporation of L-3,4-Dihydroxyphenylalanine (L-DOPA).
|
| ジャーナル |
Angew Chem Int Ed Engl
|
| Abstract |
The catechol group of 3,4-dihydroxyphenylalanine (L-DOPA) derived from L-tyrosine oxidation is a key post-translational modification (PTM) in many protein biomaterials and has potential as a bioorthogonal handle for precision protein conjugation applications such as antibody-drug conjugates. Despite this potential, indiscriminate enzymatic modification of exposed tyrosine residues or complete replacement of tyrosine using auxotrophic hosts remains the preferred method of introducing the catechol moiety into proteins, which precludes many protein engineering applications. We have developed new orthogonal translation machinery to site-specifically incorporate L-DOPA into recombinant proteins and a new fluorescent biosensor to selectively monitor L-DOPA incorporation in vivo. We show simultaneous biosynthesis and incorporation of L-DOPA and apply this translation machinery to engineer a novel metalloprotein containing a DOPA-Fe chromophore.
|
| 巻・号 |
60(27)
|
| ページ |
14811-14816
|
| 公開日 |
2021-6-25
|
| DOI |
10.1002/anie.202100579
|
| PMID |
33871147
|
| PMC |
PMC8217333
|
| MeSH |
Amino Acyl-tRNA Synthetases / chemistry
Amino Acyl-tRNA Synthetases / metabolism*
Dihydroxyphenylalanine / chemistry
Dihydroxyphenylalanine / metabolism*
Models, Molecular
Molecular Structure
|
| IF |
12.959
|
| リソース情報 |
| 遺伝子材料 |
B95. delta A (RDB13711) |
