Acta Crystallographica Section D

Biological Crystallography

Volume 69, Part 5 (May 2013)

research papers

Acta Cryst. (2013). D69, 888-900    [ doi:10.1107/S0907444913002874 ]

Structural analysis of coniferyl alcohol 9-O-methyltransferase from Linum nodiflorum reveals a novel active-site environment

S. Wolters, M. Neeb, A. Berim, J. Schulze Wischeler, M. Petersen and A. Heine

Abstract: Coniferyl alcohol 9-O-methyltransferase from Linum nodiflorum (Linaceae) catalyzes the unusual methylation of the side-chain hydroxyl group of coniferyl alcohol. The protein was heterologously expressed in Escherichia coli as a hexahistidine derivative and purified for crystallization. Diffracting crystals were obtained of the pure protein and of its selenomethionine derivative, as well as of complexes with coniferyl alcohol and with S-adenosyl-L-homocysteine together with coniferyl alcohol 9-O-methyl ether (PDB entries 4ems , 4e70 and 4evi , respectively). The X-ray structures show that the phenylpropanoid binding mode differs from other phenylpropanoid O-methyltransferases such as caffeic acid O-­methyltransferase. Moreover, the active site lacks the usually conserved and catalytic histidine residue and thus implies a different reaction mode for methylation. Site-directed mutagenesis was carried out to identify critical amino acids. The binding order of coniferyl alcohol and S-adenosyl-L-­methionine was investigated by isothermal titration calorimetry experiments.

PDB references: 4ems, 4e70 and 4evi

Keywords: small-molecule O-methyltransferases; plants; S-adenosyl-L-methionine; phenylpropanoids; isothermal titration calorimetry; Linum nodiflorum.

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Supplementary Material.

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