8Z2H

Substrate analog a010 bound form of PET-degrading cutinase mutant Cut190**SS_S176A

これはPDB形式変換不可エントリーです。

8Z2H の概要

• エントリーDOI: 10.2210/pdb8z2h/pdb
• 分子名称: Alpha/beta hydrolase family protein, 4-[2-hydroxyethyloxy(oxidanyl)phosphoryl]benzoic acid, CALCIUM ION, ... (4 entities in total)
• 機能のキーワード: protein engineering, polyesterase, metal binding, aromatic ligand, hydrolase
• 由来する生物種: Saccharomonospora viridis
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 57638.66

構造登録者

Numoto, N.,Kondo, F.,Bekker, G.J.,Liao, Z.,Yamashita, M.,Iida, A.,Ito, N.,Kamiya, N.,Oda, M. (登録日: 2024-04-12, 公開日: 2024-11-06)

主引用文献

Numoto, N.,Kondo, F.,Bekker, G.J.,Liao, Z.,Yamashita, M.,Iida, A.,Ito, N.,Kamiya, N.,Oda, M.
Structural dynamics of the Ca 2+ -regulated cutinase towards structure-based improvement of PET degradation activity.
Int.J.Biol.Macromol., 281:136597-136597, 2024

PubMed Abstract: We previously revealed the structural basis of Ca dependent regulation of a polyethylene terephthalate (PET)-degrading enzyme, Cut190, and proposed a unique reaction cycle in which the enzyme repeatedly binds and releases Ca. Here, we report crystal structures of Cut190 mutants with high thermal stability complexed with PET-like ligands that contain aromatic rings. The structural information has allowed us to perform further computational analyses using a PET-trimer bound model. Our multicanonical molecular dynamics simulations and subsequent analyses of the free energy landscapes revealed a novel intermediate form that occurs during the enzymatic reaction cycle. Furthermore, the computational analyses were used to investigate the effect of the point mutations F77L and F81L in the Ca-binding site, which showed that the former stabilizes the engaged and open forms to improve transition between the open and active forms, while the latter extremely increases the open form. Subsequent experiments showed that the F77L mutation increased the activity, while the F81L mutation decreased the activity. Our computational analysis has enabled us to explore the dynamics of Cut190 on a completely new level, providing key insights into how the balance between the various conformations influences the reaction cycle and ultimately how to improve the reaction cycle.

PubMed: 39419144
DOI: 10.1016/j.ijbiomac.2024.136597

実験手法

X-RAY DIFFRACTION (1.8 Å)