Sandbox Wabash 23 Fumarase

Benjamin Washer

Function

Fumarase C is an enzyme found in E. coli (EfumC) that is similar to the cytosolic and mitochondrial enzymes found in eukaryotic cells and is used to catalyze the hydration/dehydration reaction between L-malate and fumarate. Because of its relative simplicity, homology with important eukaryotic enzymes and important metabolic influence, fumarase has been the object of much study. However, upon analysis of fumarase, it was found that there were two "active" sites and much debate has arisen regarding which is the most influential or "true" one. This article describes the debate and the process used to determine the true active site.

Relevance

Structural highlights

The general mechanistic idea about the fumarase reaction is that at the active site, L-malate is converted to fumarate via several steps. First, the a proton is removed at the C3 by a basic residue near the active site forming an aci-carboxylate intermediate. In the second step, the -OH group on the C2 carbon leaves as an OH- ion. This is step is thought to be aided by another protonated basic group on the enzyme. However, upon X-ray crystallography analysis of the E-S complex, it was found that there were traces of L-malate at , where His is the proton transfer mediator (The red His referred to as site A and the blue His is referred to as site B). This raises the issue as to which is the the true site. It is thought that while the B site has several stabilizing residues such as , the A site is formed by residues from three of the four subunits are shown here.