1MP3

L89T VARIANT OF S. ENTERICA RmlA

1MP3 の概要

• エントリーDOI: 10.2210/pdb1mp3/pdb
• 関連するPDBエントリー: 1IIM 1IIN 1MP4 1MP5
• 分子名称: GLUCOSE-1-PHOSPHATE THYMIDYLYLTRANSFERASE, THYMIDINE-5'-TRIPHOSPHATE (2 entities in total)
• 機能のキーワード: transferase
• 由来する生物種: Salmonella enterica
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 66758.76

構造登録者

Barton, W.A.,Biggins, J.B.,Jiang, J.,Thorson, J.T.,Nikolov, D.B. (登録日: 2002-09-11, 公開日: 2002-10-09, 最終更新日: 2024-02-14)

主引用文献

Barton, W.A.,Biggins, J.B.,Jiang, J.,Thorson, J.T.,Nikolov, D.B.
Expanding pyrimidine diphosphosugar libraries via structure-based nucleotidylyltransferase engineering
Proc.Natl.Acad.Sci.USA, 99:13397-13402, 2002

PubMed Abstract: In vitro "glycorandomization" is a chemoenzymatic approach for generating diverse libraries of glycosylated biomolecules based on natural product scaffolds. This technology makes use of engineered variants of specific enzymes affecting metabolite glycosylation, particularly nucleotidylyltransferases and glycosyltransferases. To expand the repertoire of UDP/dTDP sugars readily available for glycorandomization, we now report a structure-based engineering approach to increase the diversity of alpha-d-hexopyranosyl phosphates accepted by Salmonella enterica LT2 alpha-d-glucopyranosyl phosphate thymidylyltransferase (E(p)). This article highlights the design rationale, determined substrate specificity, and structural elucidation of three "designed" mutations, illustrating both the success and unexpected outcomes from this type of approach. In addition, a single amino acid substitution in the substrate-binding pocket (L89T) was found to significantly increase the set of alpha-d-hexopyranosyl phosphates accepted by E(p) to include alpha-d-allo-, alpha-d-altro-, and alpha-d-talopyranosyl phosphate. In aggregate, our results provide valuable blueprints for altering nucleotidylyltransferase specificity by design, which is the first step toward in vitro glycorandomization.

PubMed: 12374866
DOI: 10.1073/pnas.192468299

実験手法

X-RAY DIFFRACTION (2.2 Å)