4E70

Crystal Structure Analysis of Coniferyl Alcohol 9-O-Methyltransferase from Linum Nodiflorum in Complex with Coniferyl Alcohol

Summary for 4E70

• Entry DOI: 10.2210/pdb4e70/pdb
• Descriptor: Coniferyl alcohol 9-O-methyltransferase, GLYCEROL, 4-[(1E)-3-hydroxyprop-1-en-1-yl]-2-methoxyphenol, ... (4 entities in total)
• Functional Keywords: s-adenosyl-l-methionine, small molecule o-methyltransferase, coniferyl alcohol, transferase, dimer, rossmann fold
• Biological source: Linum nodiflorum
• Total number of polymer chains: 2
• Total formula weight: 87487.67

Authors

Wolters, S.,Heine, A.,Petersen, M. (deposition date: 2012-03-16, release date: 2013-05-01, Last modification date: 2024-02-28)

Primary citation

Wolters, S.,Neeb, M.,Berim, A.,Schulze Wischeler, J.,Petersen, M.,Heine, A.
Structural analysis of coniferyl alcohol 9-O-methyltransferase from Linum nodiflorum reveals a novel active-site environment.
Acta Crystallogr.,Sect.D, 69:888-900, 2013

PubMed 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.

PubMed: 23633600
DOI: 10.1107/S0907444913002874

Experimental method

X-RAY DIFFRACTION (1.6093 Å)