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Sandbox Wabash 27 Fumarase
From Proteopedia
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==Fumarase: The Active Site Debate Answered== | ==Fumarase: The Active Site Debate Answered== | ||
<StructureSection load='1YFE' size='340' side='right' caption='Fumarase' scene=''> | <StructureSection load='1YFE' size='340' side='right' caption='Fumarase' scene=''> | ||
| - | Fumarase C (fumarate hydratase) is an enzyme found in eukaryotic cells which catalyzes the reaction between L-malate and fumarate.<ref>PMID: 9098893</ref> The catalysis proceeds via the deprotination of the C3 carbon of L-malate, which is followed by the loss of the -OH group attached to the C2 carbon; the intermediate for which is a carbanion transition state.<ref>Voet, Donald, Judith G. Voet, and Charlotte W. Pratt. Fundamentals of Biochemistry. 3rd ed. New York: Wiley, 1999. Print.</ref> In order for this catalysis to occur L-malate must be bound to fumarase. Studies have determined that fumarase has two carboxylic binding sites (site-A and site-B) which could potentially be the active site for the conversion of L-malate to fumarate. Mutations of these sites were performed in order to determine their effects on the enzymatic activity of the fumarases. The results showed that a mutation of site-B lead to no statistical difference in specific activity when compared to wild type activity, while site-A's mutation lead to a significant decrease in enzyme activity. Thus, it was determined that binding site-A is in fact the active site for the conversion of L-malate to fumarate <ref>PMID:9098893</ref>. | + | Fumarase C (fumarate hydratase) is an enzyme found in eukaryotic cells which catalyzes the reaction between L-malate and fumarate.<ref>PMID: 9098893</ref> The catalysis proceeds via the deprotination of the C3 carbon of L-malate, which is followed by the loss of the -OH group attached to the C2 carbon; the intermediate for which is a carbanion transition state.<ref>Voet, Donald, Judith G. Voet, and Charlotte W. Pratt. Fundamentals of Biochemistry. 3rd ed. New York: Wiley, 1999. Print.</ref> In order for this catalysis to occur L-malate must be bound to fumarase. Studies have determined that fumarase has two carboxylic binding sites (site-A and site-B) which could potentially be the active site for the conversion of L-malate to fumarate. Mutations of these sites were performed in order to determine their effects on the enzymatic activity of the fumarases. Specifically, the key histidines which were known to play an active role in the binding process were removed for each respective site. The results showed that a mutation of site-B lead to no statistical difference in specific activity when compared to wild type activity, while site-A's mutation lead to a significant decrease in enzyme activity. Thus, it was determined that binding site-A is in fact the active site for the conversion of L-malate to fumarate <ref>PMID:9098893</ref>. |
== Binding Site Characteristics == | == Binding Site Characteristics == | ||
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== H188N: Effects of Mutation == | == H188N: Effects of Mutation == | ||
| - | H188N is used to refer to the | + | H188N is used to refer to the mutation which replaced the H188 residue in site-A with asparagine. |
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== H129N: Effects of Mutation == | == H129N: Effects of Mutation == | ||
| - | H129N is used to refer to the | + | H129N is used to refer to the mutation which replaced the H129 rediue in site-B with asparagine. |
== Summary and Conclusions == | == Summary and Conclusions == | ||
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This is a sample scene created with SAT to <scene name="/12/3456/Sample/1">color</scene> by Group, and another to make <scene name="/12/3456/Sample/2">a transparent representation</scene> of the protein. You can make your own scenes on SAT starting from scratch or loading and editing one of these sample scenes. | This is a sample scene created with SAT to <scene name="/12/3456/Sample/1">color</scene> by Group, and another to make <scene name="/12/3456/Sample/2">a transparent representation</scene> of the protein. You can make your own scenes on SAT starting from scratch or loading and editing one of these sample scenes. | ||
Revision as of 03:45, 28 February 2016
Fumarase: The Active Site Debate Answered
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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
- ↑ Voet, Donald, Judith G. Voet, and Charlotte W. Pratt. Fundamentals of Biochemistry. 3rd ed. New York: Wiley, 1999. Print.
- ↑ 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
