Dataset Information

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

ABSTRACT: Glycosyltransferases are Nature’s versatile tools to tailor the functionalities of proteins, carbohydrates, lipids, and small molecules by transferring sugars, while C13-apocarotenoids are oxidative degradation products of carotenoids/xanthophylls that fulfil multiple physiological functions. Here, we show that C13-apocarotenoids also interact with the plant glycosyltransferase NbUGT72AY1, ultimately reducing its inherent UDP-α-D-glucose glucohydrolase activity and increasing its catalytic activity for productive substrates such as hydroxycoumarins, which are natural plant protective agents. In contrast, C13-apocarotenoids show no effect on the catalytic activity of the enzyme toward monolignol lignin precursors. Hydrogen deuterium exchange mass spectrometry revealed opposite conformational changes near the catalytic site in the presence of substrates and modulators and studies in tobacco plants confirmed increased glycosylation activity from apocarotenoid supplementation. The interaction of two plant protection mechanisms and the function of apocarotenoids as allosteric enhancers of catalytic proteins has significant implications for potential applications of these multifunctional natural products in agriculture as well as in the pharmaceutical industry.

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Escherichia Coli Nicotiana Benthamiana

SUBMITTER: Christina Ludwig

LAB HEAD: Christina Ludwig

PROVIDER: PXD035955 | Pride | 2023-03-30

REPOSITORIES: Pride

ACCESS DATA

Publications

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

Sun Guangxin G Liao Jieren J Kurze Elisabeth E Hoffmann Timothy D TD Steinchen Wieland W McGraphery Kate K Habegger Ruth R Marek Ludwig L Catici Dragana A M DAM Ludwig Christina C Jing Tingting T Hoffmann Thomas T Song Chuankui C Schwab Wilfried W

The New phytologist 20230330 5

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 glycosyl ...[more]

PMID: 36908092

Similar Datasets

Project description:As biopharmaceuticals, recombinant proteins have become indispensable tools in medicine. An increasing demand, not only in quantity but also in diversity, drives the constant development and improvement of production platforms. The N-glycosylation pattern on biopharmaceuticals plays important roles in activity, serum half-life and immunogenicity. Therefore, production platforms with tailored protein N-glycosylation are of great interest. Plant-based systems have already demonstrated their potential to produce pharmaceutically relevant recombinant proteins, although their N-glycan patterns differ from those in humans. Plants have shown great plasticity towards the manipulation of their glycosylation machinery, and some have already been glyco-engineered in order to avoid the attachment of plant-typical, putatively immunogenic sugar residues. This resulted in complex-type N-glycans with a core structure identical to the human one. Compared to humans, plants lack the ability to elongate these N-glycans with β1,4-linked galactoses and terminal sialic acids. However, these modifications, which require the activity of several mammalian enzymes, have already been achieved for Nicotiana benthamiana and the moss Physcomitrella. Here, we present the first step towards sialylation of recombinant glycoproteins in Physcomitrella, human β1,4-linked terminal N-glycan galactosylation, which was achieved by the introduction of a chimeric β1,4-galactosyltransferase (FTGT). This chimeric enzyme consists of the moss α1,4-fucosyltransferase transmembrane domain, fused to the catalytic domain of the human β1,4-galactosyltransferase. Stable FTGT expression led to the desired β1,4-galactosylation. However, additional pentoses of unknown identity were also observed. The nature of these pentoses was subsequently determined by Western blot and enzymatic digestion followed by mass spectrometric analysis and resulted in their identification as α-linked arabinoses. Since a pentosylation of β1,4-galactosylated N-glycans was reported earlier, e.g. on recombinant human erythropoietin produced in glyco-engineered Nicotiana tabacum, this phenomenon is of a more general importance for plant-based production platforms. Arabinoses, which are absent in humans, may prevent the full humanization of plant-derived products. Therefore, the identification of these pentoses as arabinoses is important as it creates the basis for their abolishment to ensure the production of safe biopharmaceuticals in plant-based systems.

Dataset Information

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

Publications

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

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure