4EVI

Crystal Structure Analysis of Coniferyl Alcohol 9-O-Methyltransferase from Linum Nodiflorum in Complex with Coniferyl Alcohol 9-Methyl Ether and S -Adenosyl-L-Homocysteine

Summary for 4EVI

• Entry DOI: 10.2210/pdb4evi/pdb
• Related: 4E70 4EMS
• Descriptor: Coniferyl alcohol 9-O-methyltransferase, S-ADENOSYL-L-HOMOCYSTEINE, 2-methoxy-4-[(1E)-3-methoxyprop-1-en-1-yl]phenol, ... (6 entities in total)
• Functional Keywords: rossmann fold, dimer, small molecule o-methyltransferase, transferase, s-adenosyl-l-methionine, coniferyl alcohol, s -adenosyl-l-homocysteine
• Biological source: Linum nodiflorum
• Total number of polymer chains: 2
• Total formula weight: 87978.20

Authors

Wolters, S.,Heine, A.,Petersen, M. (deposition date: 2012-04-26, release date: 2013-05-01, Last modification date: 2023-09-13)

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 (2.015 Å)