1HYO

CRYSTAL STRUCTURE OF FUMARYLACETOACETATE HYDROLASE COMPLEXED WITH 4-(HYDROXYMETHYLPHOSPHINOYL)-3-OXO-BUTANOIC ACID

1HYO の概要

• エントリーDOI: 10.2210/pdb1hyo/pdb
• 関連するPDBエントリー: 1QCN 1QCO 1QQJ
• 分子名称: FUMARYLACETOACETATE HYDROLASE, MAGNESIUM ION, CALCIUM ION, ... (7 entities in total)
• 機能のキーワード: beta-sandwich roll, hydrolase
• 由来する生物種: Mus musculus (house mouse)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 93259.06

構造登録者

Bateman, R.L.,Bhanumoorthy, P.,Witte, J.F.,McClard, R.W.,Grompe, M.,Timm, D.E. (登録日: 2001-01-21, 公開日: 2001-02-14, 最終更新日: 2023-08-09)

主引用文献

Bateman, R.L.,Bhanumoorthy, P.,Witte, J.F.,McClard, R.W.,Grompe, M.,Timm, D.E.
Mechanistic inferences from the crystal structure of fumarylacetoacetate hydrolase with a bound phosphorus-based inhibitor.
J.Biol.Chem., 276:15284-15291, 2001

PubMed Abstract: Fumarylacetoacetate hydrolase (FAH) catalyzes the hydrolytic cleavage of a carbon-carbon bond in fumarylacetoacetate to yield fumarate and acetoacetate as the final step of Phe and Tyr degradation. This unusual reaction is an essential human metabolic function, with loss of FAH activity causing the fatal metabolic disease hereditary tyrosinemia type I (HT1). An enzymatic mechanism involving a catalytic metal ion, a Glu/His catalytic dyad, and a charged oxyanion hole was previously proposed based on recently determined FAH crystal structures. Here we report the development and characterization of an FAH inhibitor, 4-(hydroxymethylphosphinoyl)-3-oxo-butanoic acid (HMPOBA), that competes with the physiological substrate with a K(i) of 85 microM. The crystal structure of FAH complexed with HMPOBA refined at 1.3-A resolution reveals the molecular basis for the competitive inhibition, supports the proposed formation of a tetrahedral alkoxy transition state intermediate during the FAH catalyzed reaction, and reveals a Mg(2+) bound in the enzyme's active site. The analysis of FAH structures corresponding to different catalytic states reveals significant active site side-chain motions that may also be related to catalytic function. Thus, these results advance the understanding of an essential catabolic reaction associated with a fatal metabolic disease and provide insight into the structure-based development of FAH inhibitors.

PubMed: 11154690
DOI: 10.1074/jbc.M007621200

実験手法

X-RAY DIFFRACTION (1.3 Å)