3DOH

Crystal Structure of a Thermostable Esterase

3DOH の概要

• エントリーDOI: 10.2210/pdb3doh/pdb
• 関連するPDBエントリー: 3DOI
• 分子名称: esterase, SULFATE ION (3 entities in total)
• 機能のキーワード: alpha-beta hydrolase, beta sheet, hydrolase
• 由来する生物種: Thermotoga maritima
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 87096.13

構造登録者

Levisson, M.,Sun, L.,Hendriks, S.,Dijkstra, B.W.,Van der Oost, J.,Kengen, S.W.M. (登録日: 2008-07-04, 公開日: 2009-02-17, 最終更新日: 2024-03-20)

主引用文献

Levisson, M.,Sun, L.,Hendriks, S.,Swinkels, P.,Akveld, T.,Bultema, J.B.,Barendregt, A.,van den Heuvel, R.H.H.,Dijkstra, B.W.,van der Oost, J.,Kengen, S.W.M.
Crystal structure and biochemical properties of a novel thermostable esterase containing an immunoglobulin-like domain.
J.Mol.Biol., 385:949-962, 2009

PubMed Abstract: Comparative analysis of the genome of the hyperthermophilic bacterium Thermotoga maritima revealed a hypothetical protein (EstA) with typical esterase features. The EstA protein was functionally produced in Escherichia coli and purified to homogeneity. It indeed displayed esterase activity with optima at or above 95 degrees C and at pH 8.5, with a preference for esters with short acyl chains (C2-C10). Its 2.6-A-resolution crystal structure revealed a classical alpha/beta hydrolase domain with a catalytic triad consisting of a serine, an aspartate, and a histidine. EstA is irreversibly inhibited by the organophosphate paraoxon. A 3.0-A-resolution structure confirmed that this inhibitor binds covalently to the catalytic serine residue of EstA. Remarkably, the structure also revealed the presence of an N-terminal immunoglobulin (Ig)-like domain, which is unprecedented among esterases. EstA forms a hexamer both in the crystal and in solution. Electron microscopy showed that the hexamer in solution is identical with the hexamer in the crystal, which is formed by two trimers, with the N-terminal domains facing each other. Mutational studies confirmed that residues Phe89, Phe112, Phe116, Phe246, and Trp377 affect enzyme activity. A truncated mutant of EstA, in which the Ig-like domain was removed, showed only 5% of wild-type activity, had lower thermostability, and failed to form hexamers. These data suggest that the Ig-like domain plays an important role in the enzyme multimerization and activity of EstA.

PubMed: 19013466
DOI: 10.1016/j.jmb.2008.10.075

実験手法

X-RAY DIFFRACTION (2.6 Å)