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Literature summary for 1.1.1.283 extracted from
- Structural insights into the cofactor-assisted substrate recognition of yeast methylglyoxal/isovaleraldehyde reductase Gre2 (2014), Biochim. Biophys. Acta, 1844, 1486-1492 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene gre2, recombinant expression of His6-tagged type and mutant enzymes in Escherichia coli strain BL21(DE3) | Saccharomyces cerevisiae |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| purified recombinant enzyme in apoform and in a complex with NADPH, X-ray diffraction structure determination and analysis at 2.0 A and 2.4 A, respectively | Saccharomyces cerevisiae |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| F132A/V162A | the replacement of Phe132/Val162, with Ala results in the elevation of enzymatic activity toward isovaleraldehyde, probably via enlarging the pocket entrance | Saccharomyces cerevisiae |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| isovaleraldehyde + NADPH + H+ | Saccharomyces cerevisiae | catalytic mechanism involving Ser127, Tyr165, and Lys169, overview. The carbonyl oxygen interactswith the side chain of Ser127, Tyr165 through hydrogen bonds (about 2.7 A), giving a distance of 3.0 A between the C4 atom of the nicotinamide and the carbonyl carbon of substrate | isoamyl alcohol + NADP+ | - |
? |  |
| methylglyoxal + NADPH + H+ | Saccharomyces cerevisiae | - |
(S)-lactaldehyde + NADP+ | - |
r | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Saccharomyces cerevisiae | Q12068 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration, followed by ultrafiltration | Saccharomyces cerevisiae |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| (S)-lactaldehyde + NADP+ = 2-oxopropanal + NADPH + H+ | catalytic mechanism involving Ser127, Tyr165, and Lys169, overview. In the first step, the hydroxyl groups of the Ser127 and Tyr165 residues form hydrogen bonds with the carbonyl oxygen of isovaleraldehyde, stabilizing its position. Subsequently, the hydroxyl group of Tyr165 donates a hydrogen to the carbonyl through deprotonation. Concomitantly, NADPH releases a hydrogen from the B-face of the nicotinamide ring onto the susceptible position of the carbonyl group. Thus, the carbonyl group is reduced to an alcohol group, and the isoamyl alcohol and NADP+ are produced. Upon reduction of the carbonyl group, the hydrogen bond between the side chain of Tyr165 and isoamyl alcohol is broken. With the redox state change, the conformation of NADP+ also may change, accompanied by the opening of the interdomain cleft for the release of the product | Saccharomyces cerevisiae |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (S)-lactaldehyde + NADP+ | - |
Saccharomyces cerevisiae | methylglyoxal + NADPH + H+ | - |
r | |
| isovaleraldehyde + NADPH + H+ | catalytic mechanism involving Ser127, Tyr165, and Lys169, overview. The carbonyl oxygen interactswith the side chain of Ser127, Tyr165 through hydrogen bonds (about 2.7 A), giving a distance of 3.0 A between the C4 atom of the nicotinamide and the carbonyl carbon of substrate | Saccharomyces cerevisiae | isoamyl alcohol + NADP+ | - |
? | |
| isovaleraldehyde + NADPH + H+ | catalytic mechanism involving Ser127, Tyr165, and Lys169, overview. The carbonyl oxygen interacts with the side chain of Ser127, Tyr165 through hydrogen bonds (about 2.7 A), giving a distance of 3.0 A between the C4 atom of the nicotinamide and the carbonyl carbon of substrate | Saccharomyces cerevisiae | isoamyl alcohol + NADP+ | - |
? | |
| methylglyoxal + NADPH + H+ | - |
Saccharomyces cerevisiae | (S)-lactaldehyde + NADP+ | - |
r | |
| additional information | the substrate recognition and the catalytic mechanism underlie the stereoselective reduction of Gre2. Analysis of the substrate-binding site using computational simulation and enzymatic activity assays, noticeable induced fit upon NADPH binding, overview. In Gre2, the hydrophobic residues Phe85, Tyr128 and Tyr198 combine with Phe132 and Val162 to form one funneled pocket which consists of one broad pocket entrance and one deep hydrophobic channel. The extended hydrophobic entrance of Gre2 plays a role in accommodating a wide variety of carbonyl compounds, such as diketones, aliphatic and cyclic alpha- and beta-keto esters and aldehydes.The deep hydrophobic channel prefers to identify a substrate with a linear substrate. That is why Gre2 shows high reduction activity to butanal, pentanal and 2,5-hexanedione, as well as some aldehydes | Saccharomyces cerevisiae | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homodimer | enzyme Gre2 forms a homodimer, each subunit of which contains an N-terminal Rossmann-fold domain and a variable C-terminal domain, which participates in substrate recognition. The induced fit upon binding to the cofactor NADPH makes the two domains shift toward each other, producing an interdomain cleft that better fits the substrate | Saccharomyces cerevisiae |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| Gre2 | - |
Saccharomyces cerevisiae |
| GRE2/YOL151W | - |
Saccharomyces cerevisiae |
| methylglyoxal/isovaleraldehyde reductase | - |
Saccharomyces cerevisiae |
| NADPH-dependent methylglyoxal reductase | - |
Saccharomyces cerevisiae |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
assay at | Saccharomyces cerevisiae |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
assay at | Saccharomyces cerevisiae |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NADP+ | - |
Saccharomyces cerevisiae | |
| NADPH | dependent on | Saccharomyces cerevisiae | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | the enzyme belongs to the short-chain dehydrogenase/reductase (SDR) superfamily, which includes various oxidoreductases, some isomerases and lyases | Saccharomyces cerevisiae |
| additional information | Gre2 forms a homodimer, each subunit of which contains an N-terminal Rossmann-fold domain and a variable C-terminal domain, which participates in substrate recognition. The induced fit upon binding to the cofactor NADPH makes the two domains shift toward each other, producing an interdomain cleft that better fits the substrate. The substrate-binding pocket structure determines the stringent substrate stereoselectivity for catalysis | Saccharomyces cerevisiae |
| physiological function | the Saccharomyces cerevisiae enzyme serves as a versatile enzyme that catalyzes the stereoselective reduction of a broad range of substrates including aliphatic and aromatic ketones, diketones, as well as aldehydes, using NADPH as the cofactor | Saccharomyces cerevisiae |
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Release 2023.1 (January 2023)
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