Subfunctionalization of a monolignol to a phytoalexin glucosyltransferase is accompanied by substrate inhibition

Liao, Jieren; Sun, Guangxin; Kurze, Elisabeth; Steinchen, Wieland; Hoffmann, Timothy D; Song, Chuankui; Zou, Zhiwei; Hoffmann, Thomas; Schwab, Wilfried

doi:10.1016/j.xplc.2022.100506

Cited by 7 publications

(24 citation statements)

References 82 publications

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“…The effects of scopoletin and UDP binding on the structure of NbUGT72AY1 were previously analyzed by HDX-MS (Liao et al, 2022). In the present study, NbUGT72AY1 was subjected The pattern of deuterium incorporation by GST-NbUGT72AY1 showed low deuteration in the N-terminal GST portion of the fusion protein but high deuterium exchange in the loop connecting GST to NbUGT72AY1.…”

Section: Hydrogen/deuterium Exchange Mass Spectrometry (Hdx) Disclose...mentioning

confidence: 85%

“…The effects of scopoletin and UDP binding on the structure of NbUGT72AY1 were previously analyzed by HDX‐MS (Liao et al ., 2022). In the present study, NbUGT72AY1 was subjected to HDX‐MS in the absence and in presence of scopoletin and/or effector ligands.…”

Section: Resultsmentioning

confidence: 99%

“…Recently, NbUGT72AY1 was shown to exhibit non‐Michaelis–Menten kinetics, which was interpreted as substrate inhibition (Liao et al ., 2022). To investigate the effect of apocarotenoids on the non‐Michaelis–Menten kinetics of scopoletin glucosylation, we determined the formation of scopoletin glucoside in the absence and presence of apocarotenoids by LC–MS (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…In Fragaria vesca , binding of calcium/calmodulin to an anthocyanidin UGT was shown to promote anthocyanin accumulation by reducing substrate inhibition (Peng et al ., 2016). Because substrate inhibition of NbUGT72AY1 was recently described for scopoletin, apocarotenoids might enhance glycosyltransferase activity of the enzyme by inhibiting scopoletin substrate inhibition in a manner similar to calmodulin (Liao et al ., 2022).…”

Section: Discussionmentioning

confidence: 99%

“…During a study of the monolignol/phytoalexin NbUGT72AY1 (Sun et al, 2019(Sun et al, , 2020Liao et al, 2022), we detected substrate inhibition by the acceptor-substrate scopoletin at 4 μM and discovered the enzyme's promiscuous activity for C13apocarotenoids. Detailed investigations revealed significant UDP-glucose glucosehydrolase activity of the protein, which was inhibited by apocarotenoids.…”

Section: Introductionmentioning

confidence: 99%

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Apocarotenoids are allosteric effectors of a dimeric plant glycosyltransferase involved in defense and lignin formation

et al. 2023

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Summary Glycosyltransferases are nature's versatile tools to tailor the functionalities of proteins, carbohydrates, lipids, and small molecules by transferring sugars. Prominent substrates are hydroxycoumarins such as scopoletin, which serve as natural plant protection agents. Similarly, C13‐apocarotenoids, which are oxidative degradation products of carotenoids/xanthophylls, protect plants by repelling pests and attracting pest predators. We show that C13‐apocarotenoids interact with the plant glycosyltransferase NbUGT72AY1 and induce conformational changes in the enzyme catalytic center ultimately reducing its inherent UDP‐α‐d‐glucose glucohydrolase activity and increasing its catalytic activity for productive hydroxycoumarin substrates. By contrast, C13‐apocarotenoids show no effect on the catalytic activity toward monolignol lignin precursors, which are competitive substrates. In vivo studies in tobacco plants (Nicotiana benthamiana) confirmed increased glycosylation activity upon apocarotenoid supplementation. Thus, hydroxycoumarins and apocarotenoids represent specialized damage‐associated molecular patterns, as they each provide precise information about the plant compartments damaged by pathogen attack. The molecular basis for the C13‐apocarotenoid‐mediated interplay of two plant protective mechanisms and their function as allosteric enhancers opens up potential applications of the natural products in agriculture and pharmaceutical industry.

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Section: Hydrogen/deuterium Exchange Mass Spectrometry (Hdx) Disclose...mentioning

confidence: 85%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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