3PKP

Q83S Variant of S. Enterica RmlA with dATP

3PKP の概要

• エントリーDOI: 10.2210/pdb3pkp/pdb
• 関連するPDBエントリー: 1IIM 1IIN 1MP3 1MP4 1MP5 3PKQ
• 分子名称: Glucose-1-phosphate thymidylyltransferase, 2'-DEOXYADENOSINE 5'-TRIPHOSPHATE, MAGNESIUM ION, ... (4 entities in total)
• 機能のキーワード: nucleotidylyltransferase, directed evolution, structural genomics, psi-2, protein structure initiative, center for eukaryotic structural genomics, cesg, transferase
• 由来する生物種: Salmonella enterica subsp. enterica serovar Typhimurium
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 263879.94

構造登録者

Chang, A.,Moretti, R.,Bingman, C.A.,Thorson, J.S.,Phillips Jr., G.N.,Center for Eukaryotic Structural Genomics (CESG) (登録日: 2010-11-11, 公開日: 2011-01-12, 最終更新日: 2024-02-21)

主引用文献

Moretti, R.,Chang, A.,Peltier-Pain, P.,Bingman, C.A.,Phillips, G.N.,Thorson, J.S.
Expanding the Nucleotide and Sugar 1-Phosphate Promiscuity of Nucleotidyltransferase RmlA via Directed Evolution.
J.Biol.Chem., 286:13235-13243, 2011

PubMed Abstract: Directed evolution is a valuable technique to improve enzyme activity in the absence of a priori structural knowledge, which can be typically enhanced via structure-guided strategies. In this study, a combination of both whole-gene error-prone polymerase chain reaction and site-saturation mutagenesis enabled the rapid identification of mutations that improved RmlA activity toward non-native substrates. These mutations have been shown to improve activities over 10-fold for several targeted substrates, including non-native pyrimidine- and purine-based NTPs as well as non-native D- and L-sugars (both α- and β-isomers). This study highlights the first broadly applicable high throughput sugar-1-phosphate nucleotidyltransferase screen and the first proof of concept for the directed evolution of this enzyme class toward the identification of uniquely permissive RmlA variants.

PubMed: 21317292
DOI: 10.1074/jbc.M110.206433

実験手法

X-RAY DIFFRACTION (2.6 Å)