5UD5

Crystal structure of the tRNA binding domain of Pyrrolysyl-tRNA synthetase bound to tRNA(Pyl)

5UD5 の概要

• エントリーDOI: 10.2210/pdb5ud5/pdb
• 分子名称: Pyrrolysine--tRNA ligase, RNA (70-MER), ZINC ION, ... (4 entities in total)
• 機能のキーワード: pylrs, trna, aminoacyl-trna synthetase, ligase-rna complex, ligase/rna
• 由来する生物種: Methanosarcina mazei (strain ATCC BAA-159 / DSM 3647 / Goe1 / Go1 / JCM 11833 / OCM 88); 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 71569.46

構造登録者

Suzuki, T.,Soll, D. (登録日: 2016-12-23, 公開日: 2017-10-11, 最終更新日: 2024-03-06)

主引用文献

Suzuki, T.,Miller, C.,Guo, L.T.,Ho, J.M.L.,Bryson, D.I.,Wang, Y.S.,Liu, D.R.,Soll, D.
Crystal structures reveal an elusive functional domain of pyrrolysyl-tRNA synthetase.
Nat. Chem. Biol., 13:1261-1266, 2017

PubMed Abstract: Pyrrolysyl-tRNA synthetase (PylRS) is a major tool in genetic code expansion using noncanonical amino acids, yet its structure and function are not completely understood. Here we describe the crystal structure of the previously uncharacterized essential N-terminal domain of this unique enzyme in complex with tRNA. This structure explains why PylRS remains orthogonal in a broad range of organisms, from bacteria to humans. The structure also illustrates why tRNA recognition by PylRS is anticodon independent: the anticodon does not contact the enzyme. Then, using standard microbiological culture equipment, we established a new method for laboratory evolution-a noncontinuous counterpart of the previously developed phage-assisted continuous evolution. With this method, we evolved novel PylRS variants with enhanced activity and amino acid specificity. Finally, we employed an evolved PylRS variant to determine its N-terminal domain structure and show how its mutations improve PylRS activity in the genetic encoding of a noncanonical amino acid.

PubMed: 29035363
DOI: 10.1038/nchembio.2497

実験手法

X-RAY DIFFRACTION (2.347 Å)