A catalytic triad – Lys-Asn-Asp – Is essential for the catalysis of the methyl transfer in plant cation-dependent O-methyltransferases

Brandt, Wolfgang; Manke, Kerstin; Vogt, Thomas

doi:10.1016/j.phytochem.2014.12.018

Cited by 16 publications

(14 citation statements)

References 36 publications

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“…As expected, exchange of the third type of strictly conserved residues (K157, N181, and D228, Supplementary Table SX) by amino acids of the opposite (K157D, N181Y, D228F) or even similar polarity (N181K) resulted in drastically reduced catalytic efficiencies (Figure 3c). These findings agree well with our evolutionary trace analysis (Supplementary Figure S4) and thus strongly support the hypothesis that all three residues form part of a catalytic triad required for deprotonation of the aromatic substrate prior to methyl transfer [30] …”

Section: Resultssupporting

confidence: 91%

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Altering the Regiospecificity of a CatecholO‐methyltransferase through Rational Design: Vanilloid vs. Isovanilloid Motifs in the B‐ring of Flavonoids

et al. 2022

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Rational re-design of the substrate pocket of phenylpropanoidflavonoid O-methyltransferase (PFOMT) from Mesembryanthemum crystallinum, an enzyme that selectively methylates the 3'position (= meta-position) in catechol-moieties of flavonoids to guiacol-moieties, provided the basis for the generation of variants with opposite, i. e. 4'-(para-) regioselectivity and enhanced catalytic efficiency. A double variant (Y51R/N202W) identified through a newly developed colorimetric assay efficiently modified the para-position in flavanone and flavanonol substrates, providing access to the sweetener molecule hesperetin and other rare plant flavonoids having an isovanilloid motif.

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Section: Resultssupporting

confidence: 91%

“…These findings agree well with our evolutionary trace analysis (Supplementary Figure S4) and thus strongly support the hypothesis that all three residues form part of a catalytic triad required for deprotonation of the aromatic substrate prior to methyl transfer. [30]…”

Section: Chemcatchemmentioning

confidence: 99%

Altering the Regiospecificity of a CatecholO‐methyltransferase through Rational Design: Vanilloid vs. Isovanilloid Motifs in the B‐ring of Flavonoids

et al. 2022

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“…The high conservation of amino acids results from interactions preventing their substitution and may be used for protein structural analysis. The role of one-third of the highly conserved amino acids was assigned in previous studies on various plants (Brandt et al 2015 ; Ferrer et al 2005 ; Hoffmann et al 2001 ; Kopycki et al 2008a , b ; Walker et al 2016 ). The highly conserved amino acids correspond to SbCCoAOMT-1 Gly-101, Thr-104, Ser-107, Asp-177, Asp-203, and Asn-204, which are involved in SAM or divalent metal ion binding, or Ala-178, Lys-180, and Asp-252 involved in interactions with methyl group acceptors or transmethylation products.…”

Section: Discussionmentioning

confidence: 90%

Sorghum CCoAOMT and CCoAOMT-like gene evolution, structure, expression and the role of conserved amino acids in protein activity

et al. 2018

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Sorghum is a crop plant that is grown for seeds, sucrose, forage and biofuel production. In all these applications, lignin is a superfluous component that decreases the efficiency of technological processes. Caffeoyl-coenzyme A O-methyltransferase (CCoAOMT) is an enzyme involved in monolignol synthesis that affects the efficiency of lignification and lignin composition. The sorghum genome harbors one CCoAOMT gene and six closely related CCoAOMT-like genes. The structures of four sorghum CCoAOMT-like enzymes suggest that these proteins might methylate caffeoyl coenzyme A and contribute to monolignol synthesis. In this study, two sorghum genes, CCoAOMT and one CCoAOMT-like, were found to be highly expressed in leaves, stems and immature seeds. The promoters of these genes possess clusters of transcription factor-binding sites specific for lignification, and this suggests that they are important for lignification. Phylogenetic analysis revealed that one sorghum CCoAOMT-like enzyme is closely related to ancestral cyanobacterial CCoAOMT-like proteins. The remaining CCoAOMT-like enzymes, including the one highly expressed in the leaves and stem, are closely related to CCoAOMT. Genes from these two groups possess different, evolutionarily conserved gene structures. The structure of the sorghum CCoAOMT-like protein from the ancestral clade was modeled and differences between enzymes from the two clades were analyzed. These results facilitate a better understanding of the evolution of genes involved in lignification, and provide valuable data for sorghum improvement through traditional breeding or molecular genetic techniques. The findings suggest that CCoAOMT-like genes might be recruited in lignification and raise questions of the frequency of such functional shifts.Electronic supplementary materialThe online version of this article (10.1007/s00438-018-1441-6) contains supplementary material, which is available to authorized users.

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“…S2) is debated in literature. The postulation that the conserved lysine is part of a catalytic triad is supported by the fact that the residues in question are strictly conserved in all Mg 2+ -dependent OMTs [39]. The postulation that the conserved lysine is part of a catalytic triad is supported by the fact that the residues in question are strictly conserved in all Mg 2+ -dependent OMTs [39].…”

Section: Resultsmentioning

confidence: 98%

“…It is generally accepted that the basic residue is involved in the abstraction of a proton from the hydroxyl group to be methylated; however, it is not entirely clear if the residue is essential for catalysis or only plays an accessory role [17,18,[36][37][38]. The postulation that the conserved lysine is part of a catalytic triad is supported by the fact that the residues in question are strictly conserved in all Mg 2+ -dependent OMTs [39]. We observed that the remaining activity of the lysine to alanine variants ranges from 0% to 7% of the corresponding WT activity, depending on the substrate.…”

Section: Resultsmentioning

confidence: 99%

Functional and structural characterisation of a bacterial O‐methyltransferase and factors determining regioselectivity

et al. 2017

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Mg -dependent catechol-O-methyltransferases occur in animals as well as in bacteria, fungi and plants, often with a pronounced selectivity towards one of the substrate's hydroxyl groups. Here, we show that the bacterial MxSafC exhibits excellent regioselectivity for para as well as for meta methylation, depending on the substrate's characteristics. The crystal structure of MxSafC was solved in apo and in holo form. The structure complexed with a full set of substrates clearly illustrates the plasticity of the active site region. The awareness that a wide range of factors influences the regioselectivity will aid the further development of catechol-O-methyltransferases as well as other methyltransferases as selective and efficient biocatalysts for chemical synthesis.

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A catalytic triad – Lys-Asn-Asp – Is essential for the catalysis of the methyl transfer in plant cation-dependent O-methyltransferases

Cited by 16 publications

References 36 publications

Altering the Regiospecificity of a CatecholO‐methyltransferase through Rational Design: Vanilloid vs. Isovanilloid Motifs in the B‐ring of Flavonoids

Altering the Regiospecificity of a CatecholO‐methyltransferase through Rational Design: Vanilloid vs. Isovanilloid Motifs in the B‐ring of Flavonoids

Sorghum CCoAOMT and CCoAOMT-like gene evolution, structure, expression and the role of conserved amino acids in protein activity

Functional and structural characterisation of a bacterial O‐methyltransferase and factors determining regioselectivity

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