The 1.8 Å resolution structure of hydroxycinnamoyl-coenzyme A hydratase-lyase (HCHL) fromPseudomonas fluorescens, an enzyme that catalyses the transformation of feruloyl-coenzyme A to vanillin
| Autor: | Andrzej M. Brzozowski, Derek Smith, P.M. Leonard, Andrey Lebedev, Nicholas J. Walton, Chandra S. Verma, Caroline M. Marshall, Gideon Grogan |
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| Rok vydání: | 2006 |
| Předmět: |
Models
Molecular Protein Folding Stereochemistry Coenzyme A Pseudomonas fluorescens Crystallography X-Ray Thioester Catalysis Protein Structure Secondary chemistry.chemical_compound Structural Biology Protein Structure Quaternary Enoyl-CoA Hydratase Hydro-Lyases chemistry.chemical_classification Binding Sites biology Hydrogen bond Active site Substrate (chemistry) General Medicine biology.organism_classification Lyase chemistry Benzaldehydes biology.protein Acyl Coenzyme A Oxyanion hole |
| Zdroj: | Acta Crystallographica Section D Biological Crystallography. 62:1494-1501 |
| ISSN: | 0907-4449 |
| Popis: | The crystal structure of hydroxycinnamoyl-CoA hydratase-lyase (HCHL) from Pseudomonas fluorescens AN103 has been solved to 1.8 A resolution. HCHL is a member of the crotonase superfamily and catalyses the hydration of the acyl-CoA thioester of ferulic acid [3-(4-hydroxy-3-methoxy-phenyl)prop-2-enoic acid] and the subsequent retro-aldol cleavage of the hydrated intermediate to yield vanillin (4-hydroxy-3-methoxy-benzaldehyde). The structure contains 12 molecules in the asymmetric unit, in which HCHL assumes a hexameric structure of two stacked trimers. The substrate, feruloyl-CoA, was modelled into the active site based on the structure of enoyl-CoA hydratase bound to the feruloyl-CoA-like substrate 4-(N,N-dimethylamino)-cinnamoyl-CoA (PDB code 1ey3). Feruloyl-CoA was bound in this model between helix 3 of the A subunit and helix 9 of the B subunit. A highly ordered structural water in the HCHL structure coincided with the thioester carbonyl of feruloyl-CoA in the model, suggesting that the oxyanion hole for stabilization of a thioester-derived enolate, characteristic of coenzyme-A dependent members of the crotonase superfamily, is conserved. The model also suggested that a strong hydrogen bond between the phenolic hydroxyl groups of feruloyl-CoA and BTyr239 may be an important determinant of the enzyme's ability to discriminate between the natural substrate and cinnamoyl-CoA, which is not a substrate. |
| Databáze: | OpenAIRE |
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