The Enzyme Pyruvate Decarboxylase
Image 1: Reaction catalyzed by pyruvate decarboxylase:
pyruvate + thiamine pyrophasphate (TPP) → hydroxyethyl-TPP + CO2.
Importance in Anaerobic Metabolism
Pyruvate, NADH, and ATP are the products of glycolysis. Under anaerobic conditions, pyruvate undergoes fermentation to oxidize NADH to
NAD+, so glycolysis can continue. In alcoholic fermentation, which occurs in some yeast, this is a two-step process. The first involves the Enzyme pyruvate decarboxylase (PDC). The pyruvate is decarboxylated to an acetaldehyde. This acetaldehyde then undergoes a reaction catalyzed by alcohol dehydrogenase to produce ethanol; this is the step in which the NAD+ is restored [1].
Structure
Pyruvate decarboxylase is a homotetramer. Each identical subunit consists of approximately alternating α-helices and β-sheets, and 2 domains exist within each 60kDa . This means its SCOP category is alpha and beta protein [2]. Being a homotetramer, pyruvate PDC has 4 identical that are green surrounding the ligands when the previous link is selected.
Active Site
The of PDC consists of Glu 477, Asp28, His114, and His 115 . Hydrogen bonding occurs between the substrate and Asp28, His114, and Thr73. Yeast active site residues are similar. In the catalytic step of the reaction mechanism, , shown in red, donates a proton to the pyruvate. The scene shows the close proximity of this residue to the pyruvate. The negative charge of the Glu residue following the protonation of the substrate leads to the destabilization of the pyruvate carboxylate group. Next the carboxyl group leaves. Following decarboxylation in the final step of the mechanism, release of acetaldehyde, a proton is transferred to the Glu473 residue from a cofactor. After the protonation in a concerted step, a water molecule donates a proton to the substrate while receiving a proton from Glu473. As the proton is taken from the substrate, the electrons move to form a carbonyl, which leads to the release of the acetaldehyde[3].
Regulation
PDC is regulated by substrate activation. This means that if substrate is not present in the pathway, the protein will be "off." The residue that is bound to start a cascade of events resulting in the activation of the enzyme is [4] which is highlighted in pink in the scene. This process allows the enzyme to be on when its function is necessary and off when it would not be catalyzing the reaction even if it were on.
ThDP an Important Cofactor
Thiamine diphosphate (ThDP) is an important cofactor in alpha decarboxylation reactions. The structure of ThDP is when bound to the protein. This kink brings the 4'N of ThDP in close enough proximity to C2 to it, forming a reactive ylid. Glutamic acid 51 on the other side of ThDP forms a with ThDP to increase the basicity of 4'N. In the decarboxylation reaction, of ThDP is deprotonated, and attacks C2 of the pyruvate (this structure has pyruvamide instead of pyruvate), resulting in a between ThDP and the pyruvate. This allows the ThDP to act as an electron sink for the decarboxylation reaction.
