9GTV

Crystal structure of RamR with Tyr59 replaced with para-boronophenylalanine (boronate form)

これはPDB形式変換不可エントリーです。

9GTV の概要

• エントリーDOI: 10.2210/pdb9gtv/pdb
• 分子名称: Transcriptional regulator RamR (1 entity in total)
• 機能のキーワード: artificial enzyme, tetr family, boron designer enzyme, noncanonical amino acid, para-boronophenylalanine, boron acid catalysis, dna binding protein
• 由来する生物種: Salmonella enterica subsp. enterica serovar Typhimurium
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 92440.26

構造登録者

Longwitz, L.,Brouwer, B.,Thunnissen, A.M.W.H.,Roelfes, G. (登録日: 2024-09-18, 公開日: 2024-12-25, 最終更新日: 2025-01-08)

主引用文献

Longwitz, L.,Kamer, M.D.,Brouwer, B.,Thunnissen, A.W.H.,Roelfes, G.
Boron Designer Enzyme with a Hybrid Catalytic Dyad.
Acs Catalysis, 14:18469-18476, 2024

PubMed Abstract: Genetically encoded noncanonical amino acids can introduce new-to-nature activation modes into enzymes. While these amino acids can act as catalysts on their own due to their inherent chemical properties, interactions with adjacent residues in an enzyme, such as those present in natural catalytic dyads or triads, unlock a higher potential for designer enzymes. We incorporated a boron-containing amino acid into the protein scaffold RamR to create an active enzyme for the kinetic resolution of α-hydroxythioesters. We found that a closely positioned lysine residue is crucial for the catalytic activity of the designer enzyme by forming a hybrid catalytic dyad with the boronic acid residue. The enzyme is capable of resolving differently substituted α-hydroxythioesters with good selectivities. High-resolution mass spectrometry, B NMR spectroscopy, and crystal structure analysis of the designer enzyme gave insight into the three steps of the mechanism (substrate binding, hydroxide transfer, product release). Mutations of a residue around the catalytic dyad led to a variant of the enzyme with 2-fold improvement of catalytic activity and selectivity.

PubMed: 39722884
DOI: 10.1021/acscatal.4c06052

実験手法

X-RAY DIFFRACTION (2.78 Å)