5AN7

Structure of the engineered retro-aldolase RA95.5-8F with a bound 1,3-diketone inhibitor

5AN7 の概要

• エントリーDOI: 10.2210/pdb5an7/pdb
• 関連するPDBエントリー: 5AOU
• 分子名称: RA95.5-8F, PHOSPHATE ION, (2E)-1-(6-methoxynaphthalen-2-yl)but-2-en-1-one, ... (4 entities in total)
• 機能のキーワード: lyase, retro-aldolase, protein engineering, enzyme design, directed evolution
• 由来する生物種: SULFOLOBUS SOLFATARICUS
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 30352.65

構造登録者

Obexer, R.,Mittl, P.R.E.,Hilvert, D. (登録日: 2015-09-04, 公開日: 2016-08-17, 最終更新日: 2024-11-06)

主引用文献

Obexer, R.,Godina, A.,Garrabou, X.,Mittl, P.R.,Baker, D.,Griffiths, A.D.,Hilvert, D.
Emergence of a catalytic tetrad during evolution of a highly active artificial aldolase.
Nat Chem, 9:50-56, 2017

PubMed Abstract: Designing catalysts that achieve the rates and selectivities of natural enzymes is a long-standing goal in protein chemistry. Here, we show that an ultrahigh-throughput droplet-based microfluidic screening platform can be used to improve a previously optimized artificial aldolase by an additional factor of 30 to give a >10 rate enhancement that rivals the efficiency of class I aldolases. The resulting enzyme catalyses a reversible aldol reaction with high stereoselectivity and tolerates a broad range of substrates. Biochemical and structural studies show that catalysis depends on a Lys-Tyr-Asn-Tyr tetrad that emerged adjacent to a computationally designed hydrophobic pocket during directed evolution. This constellation of residues is poised to activate the substrate by Schiff base formation, promote mechanistically important proton transfers and stabilize multiple transition states along a complex reaction coordinate. The emergence of such a sophisticated catalytic centre shows that there is nothing magical about the catalytic activities or mechanisms of naturally occurring enzymes, or the evolutionary process that gave rise to them.

PubMed: 27995916
DOI: 10.1038/nchem.2596

実験手法

X-RAY DIFFRACTION (1.1 Å)