2HZY

Mouse fumarylacetoacetate hydrolase complexes with a transition-state mimic of the complete substrate

Summary for 2HZY

• Entry DOI: 10.2210/pdb2hzy/pdb
• Related: 1HYO 1QCN 1QCO 1QQJ
• Descriptor: Fumarylacetoacetase, SODIUM ION, MANGANESE (II) ION, ... (7 entities in total)
• Functional Keywords: transition-state mimicking complex, hydrolase
• Biological source: Mus musculus (house mouse)
• Total number of polymer chains: 2
• Total formula weight: 93852.45

Authors

Hurley, T.D.,Timm, D.E. (deposition date: 2006-08-09, release date: 2006-08-22, Last modification date: 2023-08-30)

Primary citation

Bateman, R.L.,Ashworth, J.,Witte, J.F.,Baker, L.J.,Bhanumoorthy, P.,Timm, D.E.,Hurley, T.D.,Grompe, M.,McClard, R.W.
Slow-onset inhibition of fumarylacetoacetate hydrolase by phosphinate mimics of the tetrahedral intermediate: kinetics, crystal structure and pharmacokinetics.
Biochem.J., 402:251-260, 2007

PubMed Abstract: FAH (fumarylacetoacetate hydrolase) catalyses the final step of tyrosine catabolism to produce fumarate and acetoacetate. HT1 (hereditary tyrosinaemia type 1) results from deficiency of this enzyme. Previously, we prepared a partial mimic of the putative tetrahedral intermediate in the reaction catalysed by FAH co-crystallized with the enzyme to reveal details of the mechanism [Bateman, Bhanumoorthy, Witte, McClard, Grompe and Timm (2001) J. Biol. Chem. 276, 15284-15291]. We have now successfully synthesized complete mimics CEHPOBA {4-[(2-carboxyethyl)-hydroxyphosphinyl]-3-oxobutyrate} and COPHPAA {3-[(3-carboxy-2-oxopropyl)hydroxyphosphinyl]acrylate}, which inhibit FAH in slow-onset tight-binding mode with K(i) values of 41 and 12 nM respectively. A high-resolution (1.35 A; 1 A=0.1 nm) crystal structure of the FAH.CEHPOBA complex was solved to reveal the affinity determinants for these compounds and to provide further insight into the mechanism of FAH catalysis. These compounds are active in vivo, and CEHPOBA demonstrated a notable dose-dependent increase in SA (succinylacetone; a metabolite seen in patients with HT1) in mouse serum after repeated injections, and, following a single injection (1 mumol/g; intraperitoneal), only a modest regain of FAH enzyme activity was detected in liver protein isolates after 24 h. These potent inhibitors provide a means to chemically phenocopy the metabolic defects of either HT1 or FAH knockout mice and promise future pharmacological utility for hepatocyte transplantation.

PubMed: 17064256
DOI: 10.1042/BJ20060961

Experimental method

X-RAY DIFFRACTION (1.35 Å)