2J5I

Crystal Structure of Hydroxycinnamoyl-CoA Hydratase-Lyase

Summary for 2J5I

• Entry DOI: 10.2210/pdb2j5i/pdb
• Descriptor: P-HYDROXYCINNAMOYL COA HYDRATASE/LYASE, ... (4 entities in total)
• Functional Keywords: lyase, vanillin, aldolase, crotonase, hydratase, coenzyme-a
• Biological source: PSEUDOMONAS FLUORESCENS; More
• Total number of polymer chains: 12
• Total formula weight: 372502.40

Authors

Leonard, P.M.,Brzozowski, A.M.,Lebedev, A.,Marshall, C.M.,Smith, D.J.,Verma, C.S.,Walton, N.J.,Grogan, G. (deposition date: 2006-09-18, release date: 2006-12-06, Last modification date: 2023-12-13)

Primary citation

Leonard, P.M.,Brzozowski, A.M.,Lebedev, A.,Marshall, C.M.,Smith, D.J.,Verma, C.S.,Walton, N.J.,Grogan, G.
The 1.8 A Resolution Structure of Hydroxycinnamoyl- Coenzyme a Hydratase-Lyase (Hchl) from Pseudomonas Fluorescens, an Enzyme that Catalyses the Transformation of Feruloyl-Coenzyme a to Vanillin.
Acta Crystallogr.,Sect.D, 62:1494-, 2006

PubMed Abstract: 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.

PubMed: 17139085
DOI: 10.1107/S0907444906039199

Experimental method

X-RAY DIFFRACTION (1.8 Å)