Fumarase C is the stable fumarase found in E. coli acting as a "non-iron-containing enzyme" found in eukaryotic cells. Fumarase is a vital enzyme in eukaryotic cells because it has the ability to catalyze dehydration and hydration reactions between L-malate and fumarate,[1] a reaction found in the Krebs Cycle. This leaves a particular importance on the active site of fumarase, which dictates the catalytic activity of the enzyme. The first stage of the of the catalytic process is the removal of a proton from the C3 position of L-malate followed by the a deprotanation and removal of an –OH group from C2 resulting in a water molecule being formed. The presence or lack thereof of these processes is a key indicator for catalytic activity and will be used in determining the true active site of Fumarase C.
Active Site Debate
| Sample
| Site Linked to
| Average Activity (μ/mL)
| Specific Activity (μ/mg)
|
| Wild Type
| NA
| 4920
| 116
|
| H129N
| B Site
| 2080
| 143
|
| H188N
| A Site
| 10
| 0.6
|
Structure of Active Site
References
- ↑ Weaver T, Lees M, Banaszak L. Mutations of fumarase that distinguish between the active site and a nearby dicarboxylic acid binding site. Protein Sci. 1997 Apr;6(4):834-42. PMID:9098893
