Enolase

spinqualitylabelsall models Yeast enolase dimer complex with phosphoenolpyruvate and phosphoglycerate, 1one Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Enolase is an enzyme that catalyzes a reaction of glycolysis. Glycolysis converts glucose into two 3-carbon molecules called pyruvate. The energy released during glycolysis is used to make ATP. Enolase is used to convert 2-phosphoglycerate (2PG) to phosphoenolpyruvate (PEP) in the 9th reaction of glycolysis: it is a reversible dehydration reaction.. Enolase is expressed abundantly in most cells and has been proven useful as a model to study mechanisms of enzyme action and structural analysis . As with the reaction below, Enolase must have a divalent metal cation present to activate or deactivate the enzyme. The best cofactor would be Mg2+, but many, including Zn2+, Mn2+ and Co2+ can be used. The metal ion works by binding to the enzyme at the active site and producing a conformational change. This makes it possible for the substrate (2-PGA) to bind at the Enolase active site. Once this happens, a second metal ion comes in and binds to the enzyme to activate the Enolase catalytic ability. Structure The secondary structure of enolase contains both alpha helices and beta sheets. The beta sheets are mainly parallel. As shown in the figure, enolase has about 36 alpha helices and 22 beta sheets (18 alpha helices and 11 beta sheets per domain). Enolase consists of two domains. Structural Clasification of Proteins (SCOP) Enolase is in the alpha and beta proteins class and has a fold of TIM beta/alpha-barrel. It comes from the Superfamily on Enolase C-terminal domain-like and is in the enolase family. Mechanism The mechanism of 2PG to PEP using enolase. The active site of enolase as shown, involves Lys 345, Lys 396, Glu 168, Glu 211, and His 159. Enolase forms a complex with two Mg 2+'s at its active site. The substrate, 2PG, binds to the two Mg2+'s, Glu 211, and Lys 345. The Mg 2+ then forms a bond at the deprotonated carboxylic acid on the 1'C to connect it with enolase. It also is connects to Glu 211 and Lys 345. Glu 211 makes a hydrogen bond with the alcohol group on the 3'C. Lys 345 deprotonates the 2'C and then the 2'C forms an alkene with the 1'C which then moves the electrons forming the ketone onto the oxygen making it have a negative charge. The other oxygen, which already has a negative charge, then moves its electron to form a ketone with the 1'C. The electrons that made up the alkene between the 1'C adn 2'C then moves to form an alkene between the 2'C and 3'C. This breaks the bond with the alcohol on the 3'C which deprotonates Glu 211 on enolase to form H2O. Then the new molecule is released from enolase as PEP. PEP then goes on through another step in glycolysis to create pyruvate. Fluoride ions inhibits glycolysis by forming a bond with Mg 2+ thus blocks the substrate (2PG) from binding to the active site of enolase. Kinetics V vs. [PGA]; PGA is 2PG, the top curve has [Mg2+] of 10^-3 M and the bottom curve has [Mg2+] of 106-2 M Since Mg2+ is essential for binding the substrate, 2-PG, it is also needed at a specific quality in order to have a good rate, or velocity. The graph shows the V vs. [PGA], in which PGA is 2-PG, with two different concentrations of Mg2+. The upper curve, which also has greater Vmax, has an Mg2+ concentration of 10^-3 M while the lower curve, which has a lower Vmax, has an Mg2+ concentration of 10^-2 M. The Km is also larger the upper curve making the higher [Mg2+] more desirable. Regulation Enolase is found on the surface of a variety of eukaryotic cells as a strong plamingoen-binding receptor and on the surface of hematopietic celss such as monocytes, T cells and B cells, neuronal celss and endothelial cells. Enolase in muscle can bind other glycolytic enzymes, such as phosphoglycerate mutase, muscle creatine kinase, pyruvate kinase, and muscle troponin, with high affinity. This suggests that they make a functional glycolytic segment in the muscle where ATP production is required in order for the muscle to contract. Myc-binding protein (MBP-1) is similar to the a-enolse structure and is found in the nucleus as a DNA-binding protein. Enolase is regulated by the concentration of Mg2+ and the previous steps of glycolysis. Other interesting information Enolase is present in all tissues and organisms with the ability to do glycolysis or fermentation. Recent studies have Enolase concentration samples in order to determine certain conditions and their severity. For instance, high concentrations of Enolase in cerebrospinal fluid (CSF) are more strongly associated with astrocytoma than other enzymes like aldolase, pyruvate kinase, and creatine kinase. High concentrations of Enolase in the CSF are also linked to the fastest rate of tumor growth and increased chances of heart attack or stroke. Enolase can be competitively inhibited by fluoride for the substrate 2-PGA. In drinking water with added fluorination, oral bacteria Enolase activity is inhibited without harmed humans. This works to prevent cavities. 3D structures of enolase Update June 2012 Enolase 3dip, 2qgy – ENO – unidentified 1oep - TbENO – Trypanosoma brucei 2ptw – TbENO (mutant) 2ptx - TbENO (mutant) + sulfate 2pty - TbENO (mutant) + PEP 2ptz, 2pu0, 2pu1 - TbENO (mutant) + PAH 2pa6 – ENO – Methanocaldococcus jannaschii 1w6t – ENO – Streptococcus pneumoniae 1iyx – ENO – Enterococcus hirae 1pdy, 1pdz – ENO – European lobster 3qn3 – ENO – Campylobacter jejuni 3tqp – ENO – Coxiella burnetii 3ugv – ENO – Alpha proteobacterium Enolase 1 3b97, 2psn – hENO1 - human 3otr – ENO1 – Toxoplasma gondii 3qtp – ENO1 – Entamoeba histolytica 1e9i - EcENO1 – Escherichia coli 2xgz, 2xh0, 2xh2, 2xh4, 2xh7 – EcENO1 residues 2-437 (mutant) 3uj2 – ENO1 – Anaerostipes caccae 3h8a, 2fym – EcENO1 + RNase E 3enl, 4enl – yENO1 - yeast 2al1, 2al2, 1ebh – yENO1 + Mg 1p43, 1p48 – yENO1 (mutant) 1ebg, 1els - yENO1 + PAH 1l8p - yENO1 (mutant) + PAH 2one, 1one – yENO1 + PEP + phosphoglycerate 5enl, 7enl - yENO1 + phosphoglycerate 1nel, 6enl – yENO1 + inhibitor Enolase 2 1te6 – hENO2 2akm, 2akz – hENO2 + inhibitor Enolase 3 2xsx - hENO3 2, 3-diketo-5-methylthiopentyl-1-phosphate enolase 2zvi – DK-MTP-1-P ENO – Bacillus subtilis 2oej - BkDK-MTP-1-P ENO + Pi – Geobacillus kaustophilus 2oek, 2oel - BkDK-MTP-1-P ENO + ion 2oem - BkDK-MTP-1-P ENO + Mg + phosphate derivative Additional Resources For additional information, see: Carbohydrate Metabolism

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