8KE6

PylRS C-terminus domain mutant bound with L-3-chlorophenylalanine and AMPNP

8KE6 の概要

• エントリーDOI: 10.2210/pdb8ke6/pdb
• 分子名称: Pyrrolysine--tRNA ligase, PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER, MAGNESIUM ION, ... (5 entities in total)
• 機能のキーワード: trna synthetase, ligase
• 由来する生物種: Methanosarcina mazei
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 32754.04

構造登録者

Weng, J.H.,Tsai, M.D.,Wang, Y.S. (登録日: 2023-08-11, 公開日: 2023-11-01, 最終更新日: 2023-11-15)

主引用文献

Jiang, H.K.,Weng, J.H.,Wang, Y.H.,Tsou, J.C.,Chen, P.J.,Ko, A.A.,Soll, D.,Tsai, M.D.,Wang, Y.S.
Rational design of the genetic code expansion toolkit for in vivo encoding of D-amino acids.
Front Genet, 14:1277489-1277489, 2023

PubMed Abstract: Once thought to be non-naturally occurring, D-amino acids (DAAs) have in recent years been revealed to play a wide range of physiological roles across the tree of life, including in human systems. Synthetic biologists have since exploited DAAs' unique biophysical properties to generate peptides and proteins with novel or enhanced functions. However, while peptides and small proteins containing DAAs can be efficiently prepared , producing large-sized heterochiral proteins poses as a major challenge mainly due to absence of pre-existing DAA translational machinery and presence of endogenous chiral discriminators. Based on our previous work demonstrating pyrrolysyl-tRNA synthetase's (PylRS') remarkable substrate polyspecificity, this work attempts to increase PylRS' ability in directly charging tRNA with D-phenylalanine analogs (DFAs). We here report a novel, polyspecific PylRS mutant, DFRS2, capable of incorporating DFAs into proteins via ribosomal synthesis . To validate its utility, translational DAA substitution were performed in superfolder green fluorescent protein and human heavy chain ferritin, successfully altering both proteins' physiochemical properties. Furthermore, aminoacylation kinetic assays further demonstrated aminoacylation of DFAs by DFRS2 .

PubMed: 37904728
DOI: 10.3389/fgene.2023.1277489

実験手法

X-RAY DIFFRACTION (1.89570853979 Å)