Purification and characterisation of epithiospecifier protein from <i>Brassica napus</i>: enzymic intramolecular sulphur addition within alkenyl thiohydroximates derived from alkenyl glucosinolate hydrolysis

Foo, Hooi Ling; Grønning, Line M.; Goodenough, Lucy; Bones, Atle M.; Danielsen, Brit-Eli; Whiting, Don A.; Rossiter, John T.

doi:10.1016/s0014-5793(00)01176-5

Cited by 106 publications

(99 citation statements)

References 17 publications

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“…On the other hand, AtNSP2 does not require a specific myrosinase. This has also been described for the cooperative action between ESP and myrosinases (45,51,52). In our experiments AtNSP2 was able to act in conjunction with plant myrosinases from three different Brassicaceae species (i.e.…”

Section: Identification Of Genes With Sequence Similarity To Esp In Tsupporting

confidence: 83%

Nitrile-specifier Proteins Involved in Glucosinolate Hydrolysis in Arabidopsis thaliana

Kissen

Bones

2009

Journal of Biological Chemistry

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124

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Glucosinolates are plant secondary metabolites present in Brassicaceae plants such as the model plant Arabidopsis thaliana. Intact glucosinolates are believed to be biologically inactive, whereas degradation products after hydrolysis have multiple roles in growth regulation and defense. The degradation of glucosinolates is catalyzed by thioglucosidases called myrosinases and leads by default to the formation of isothiocyanates. The interaction of a protein called epithiospecifier protein (ESP) with myrosinase diverts the reaction toward the production of epithionitriles or nitriles depending on the glucosinolate structure. Here we report the identification of a new group of nitrile-specifier proteins (AtNSPs) in A. thaliana able to generate nitriles in conjunction with myrosinase and a more detailed characterization of one member (AtNSP2). Recombinant AtNSP2 expressed in Escherichia coli was used to test its impact on the outcome of glucosinolate hydrolysis using a gas chromatography-mass spectrometry approach. AtNSP proteins share 30 -45% sequence homology with A. thaliana ESP. Although AtESP and AtNSP proteins can switch myrosinase-catalyzed degradation of 2-propenylglucosinolate from isothiocyanate to nitrile, only AtESP generates the corresponding epithionitrile. Using the aromatic benzylglucosinolate, recombinant AtNSP2 is also able to direct product formation to the nitrile. Analysis of glucosinolate hydrolysis profiles of transgenic A. thaliana plants overexpressing AtNSP2 confirms its nitrile-specifier activity in planta. In silico expression analysis reveals distinctive expression patterns of AtNSPs, which supports a biological role for these proteins. In conclusion, we show that AtNSPs belonging to a new family of A. thaliana proteins structurally related to AtESP divert product formation from myrosinasecatalyzed glucosinolate hydrolysis and, thereby, likely affect the biological consequences of glucosinolate degradation. We discuss similarities and properties of AtNSPs and related proteins and the biological implications.Brassicaceae plants such as oilseed rape (Brassica napus), turnip (Brassica rapa), and white mustard (Sinapis alba) as well as the model plant Arabidopsis thaliana contain a group of secondary metabolites known as glucosinolates (GSLs) 2

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Section: Identification Of Genes With Sequence Similarity To Esp In Tsupporting

confidence: 83%

Nitrile-specifier Proteins Involved in Glucosinolate Hydrolysis in Arabidopsis thaliana

Kissen

Bones

2009

Journal of Biological Chemistry

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124

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“…Nitriles are the major degradation products under acidic or alkaline conditions and after inactivation of myrosinase. In the presence of the epithiospecifier protein (ESP) and Fe 2+ ions, myrosinase-catalysed hydrolysis of alkenyl GLSs is directed towards epithionitrile formation [153]. In consequence, different processing and storage conditions of the vegetables, e.g.…”

Section: Liberationmentioning

confidence: 99%

Glucosinolates inBrassicavegetables: The influence of the food supply chain on intake, bioavailability and human health

Verkerk¹,

Schreiner

Krumbein

et al. 2009

Molecular Nutrition Food Res

507

415

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Glucosinolates (GLSs) are found in Brassica vegetables. Examples of these sources include cabbage, Brussels sprouts, broccoli, cauliflower and various root vegetables (e.g. radish and turnip). A number of epidemiological studies have identified an inverse association between consumption of these vegetables and the risk of colon and rectal cancer. Animal studies have shown changes in enzyme activities and DNA damage resulting from consumption of Brassica vegetables or isothiocyanates, the breakdown products (BDP) of GLSs in the body. Mechanistic studies have begun to identify the ways in which the compounds may exert their protective action but the relevance of these studies to protective effects in the human alimentary tract is as yet unproven. In vitro studies with a number of specific isothiocyanates have suggested mechanisms that might be the basis of their chemoprotective effects. The concentration and composition of the GLSs in different plants, but also within a plant (e.g. in the seeds, roots or leaves), can vary greatly and also changes during plant development. Furthermore, the effects of various factors in the supply chain of Brassica vegetables including breeding, cultivation, storage and processing on intake and bioavailability of GLSs are extensively discussed in this paper.

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“…Glucosinolates such as progoitrin, sinigrin, gluconapin and glucobrassicanapin can give rise to cyanoepithioalkanes (epithionitriles). The exact mechanism behind the formation of epithionitriles from these glucosinolates is not known, but depends on the presence of a protein known as epithiospecifierprotein (ESP) (MacLeod and Rossiter, 1985;Foo et al, 2000;Zabala et al, 2005;Bones and Rossiter, 2006). The breakdown products resulting from glucosinolate hydrolysis represent the 'defence-active' components and the dual functioning of glucosinolates and myrosinases coming into contact upon tissue disruption is designated as the glucosinolate-myrosinase defence system.…”

Section: The Glucosinolate-myrosinase Defence Systemmentioning

confidence: 99%

Defence mechanisms of Brassicaceae: implications for plant-insect interactions and potential for integrated pest management. A review

Ahuja

Rohloff

Bones

2010

Agron. Sustain. Dev.

203

153

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-Brassica crops are grown worldwide for oil, food and feed purposes, and constitute a significant economic value due to their nutritional, medicinal, bioindustrial, biocontrol and crop rotation properties. Insect pests cause enormous yield and economic losses in Brassica crop production every year, and are a threat to global agriculture. In order to overcome these insect pests, Brassica species themselves use multiple defence mechanisms, which can be constitutive, inducible, induced, direct or indirect depending upon the insect or the degree of insect attack. Firstly, we give an overview of different Brassica species with the main focus on cultivated brassicas. Secondly, we describe insect pests that attack brassicas. Thirdly, we address multiple defence mechanisms, with the main focus on phytoalexins, sulphur, glucosinolates, the glucosinolate-myrosinase system and their breakdown products. In order to develop pest control strategies, it is important to study the chemical ecology, and insect behaviour. We review studies on oviposition regulation, multitrophic interactions involving feeding and host selection behaviour of parasitoids and predators of herbivores on brassicas. Regarding oviposition and trophic interactions, we outline insect oviposition behaviour, the importance of chemical stimulation, oviposition-deterring pheromones, glucosinolates, isothiocyanates, nitriles, and phytoalexins and their importance towards pest management. Finally, we review brassicas as cover and trap crops, and as biocontrol, biofumigant and biocidal agents against insects and pathogens. Again, we emphasise glucosinolates, their breakdown products, and plant volatile compounds as key components in these processes, which have been considered beneficial in the past and hold great prospects for the future with respect to an integrated pest management.Brassicas / insect pests / chemical ecology / trophic levels / glucosinolates / isothiocyanates / defence mechanisms / biocontrol / trap crops / integrated pest management

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Purification and characterisation of epithiospecifier protein from Brassica napus: enzymic intramolecular sulphur addition within alkenyl thiohydroximates derived from alkenyl glucosinolate hydrolysis

Cited by 106 publications

References 17 publications

Nitrile-specifier Proteins Involved in Glucosinolate Hydrolysis in Arabidopsis thaliana

Nitrile-specifier Proteins Involved in Glucosinolate Hydrolysis in Arabidopsis thaliana

Glucosinolates inBrassicavegetables: The influence of the food supply chain on intake, bioavailability and human health

Defence mechanisms of Brassicaceae: implications for plant-insect interactions and potential for integrated pest management. A review

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