5N5V

Structure of p-boronophenylalanyl tRNA synthetase - apo form

5N5V の概要

• エントリーDOI: 10.2210/pdb5n5v/pdb
• 関連するPDBエントリー: 5n5u
• 分子名称: Tyrosine--tRNA ligase, CHLORIDE ION (3 entities in total)
• 機能のキーワード: aminoacylation, ligase, non-natural amino acid, synthetic biology
• 由来する生物種: Methanocaldococcus jannaschii (strain ATCC 43067 / DSM 2661 / JAL-1 / JCM 10045 / NBRC 100440) (Methanococcus jannaschii)
• 細胞内の位置: Cytoplasm: Q57834
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 287991.14

構造登録者

Schiefner, A.,Skerra, A. (登録日: 2017-02-14, 公開日: 2018-02-14, 最終更新日: 2024-01-17)

主引用文献

Schiefner, A.,Nastle, L.,Landgraf, M.,Reichert, A.J.,Skerra, A.
Structural Basis for the Specific Cotranslational Incorporation of p-Boronophenylalanine into Biosynthetic Proteins.
Biochemistry, 57:2597-2600, 2018

PubMed Abstract: The site-specific incorporation of the non-natural amino acid p-boronophenylalanine (Bpa) into recombinant proteins enables the development of novel carbohydrate-binding functions as well as bioorthogonal chemical modification. To this end, Bpa is genetically encoded by an amber stop codon and cotranslationally inserted into the recombinant polypeptide chain at the ribosome by means of an artificial aminoacyl-tRNA synthetase (aaRS) in combination with a compatible suppressor tRNA. We describe the crystal structure of an aaRS specific for Bpa, which had been engineered on the basis of the TyrRS from Methanocaldococcus jannaschii, in complex with both Bpa and AMP. The substrates are bound in an orientation resembling the aminoacyl-AMP mixed anhydride intermediate and engaged in a network of four hydrogen bonds that allows specific recognition of the boronate moiety by the aaRS. The key determinant of this interaction is the coplanar alignment of its Glu162 carboxylate group with Bpa, which results in a double hydrogen bond with the boronic acid substituent. Our structural study elucidates how a small set of five side chain exchanges within the TyrRS active site can switch its substrate specificity to the hydrophilic amino acid Bpa, thus stimulating the reprogramming of other aaRS to recruit useful non-natural amino acids for next-generation protein engineering.

PubMed: 29668275
DOI: 10.1021/acs.biochem.8b00171

実験手法

X-RAY DIFFRACTION (2.3 Å)