4LCL

Simvastatin Synthase (LOVD), from Aspergillus Terreus, LovD6 mutant (simh6208)

4LCL の概要

• エントリーDOI: 10.2210/pdb4lcl/pdb
• 関連するPDBエントリー: 3HL9 3HLB 3HLC 3HLD 3HLE 3HLF 3HLG 4LCM
• 分子名称: Transesterase, ISOPROPYL ALCOHOL (3 entities in total)
• 機能のキーワード: laboratory-directed evolution, transesterase, transferase
• 由来する生物種: Aspergillus terreus
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 96908.46

構造登録者

Gao, X.,Sawaya, M.R.,Yeates, T.O.,Tang, Y. (登録日: 2013-06-21, 公開日: 2014-04-02, 最終更新日: 2023-09-20)

主引用文献

Jimenez-Oses, G.,Osuna, S.,Gao, X.,Sawaya, M.R.,Gilson, L.,Collier, S.J.,Huisman, G.W.,Yeates, T.O.,Tang, Y.,Houk, K.N.
The role of distant mutations and allosteric regulation on LovD active site dynamics.
Nat.Chem.Biol., 10:431-436, 2014

PubMed Abstract: Natural enzymes have evolved to perform their cellular functions under complex selective pressures, which often require their catalytic activities to be regulated by other proteins. We contrasted a natural enzyme, LovD, which acts on a protein-bound (LovF) acyl substrate, with a laboratory-generated variant that was transformed by directed evolution to accept instead a small free acyl thioester and no longer requires the acyl carrier protein. The resulting 29-mutant variant is 1,000-fold more efficient in the synthesis of the drug simvastatin than the wild-type LovD. This is to our knowledge the first nonpatent report of the enzyme currently used for the manufacture of simvastatin as well as the intermediate evolved variants. Crystal structures and microsecond-scale molecular dynamics simulations revealed the mechanism by which the laboratory-generated mutations free LovD from dependence on protein-protein interactions. Mutations markedly altered conformational dynamics of the catalytic residues, obviating the need for allosteric modulation by the acyl carrier LovF.

PubMed: 24727900
DOI: 10.1038/nchembio.1503

実験手法

X-RAY DIFFRACTION (1.8 Å)