By the same authors

From the same journal

From the same journal

Roles for Stress-inducible Lambda Glutathione Transferases in Flavonoid Metabolism in Plants as Identified by Ligand Fishing

Research output: Contribution to journalArticle

Standard

Roles for Stress-inducible Lambda Glutathione Transferases in Flavonoid Metabolism in Plants as Identified by Ligand Fishing. / Dixon, David P.; Edwards, Robert.

In: Journal of Biological Chemistry, Vol. 285, No. 47, 19.11.2010, p. 36322-36329.

Research output: Contribution to journalArticle

Harvard

Dixon, DP & Edwards, R 2010, 'Roles for Stress-inducible Lambda Glutathione Transferases in Flavonoid Metabolism in Plants as Identified by Ligand Fishing', Journal of Biological Chemistry, vol. 285, no. 47, pp. 36322-36329. https://doi.org/10.1074/jbc.M110.164806

APA

Dixon, D. P., & Edwards, R. (2010). Roles for Stress-inducible Lambda Glutathione Transferases in Flavonoid Metabolism in Plants as Identified by Ligand Fishing. Journal of Biological Chemistry, 285(47), 36322-36329. https://doi.org/10.1074/jbc.M110.164806

Vancouver

Dixon DP, Edwards R. Roles for Stress-inducible Lambda Glutathione Transferases in Flavonoid Metabolism in Plants as Identified by Ligand Fishing. Journal of Biological Chemistry. 2010 Nov 19;285(47):36322-36329. https://doi.org/10.1074/jbc.M110.164806

Author

Dixon, David P. ; Edwards, Robert. / Roles for Stress-inducible Lambda Glutathione Transferases in Flavonoid Metabolism in Plants as Identified by Ligand Fishing. In: Journal of Biological Chemistry. 2010 ; Vol. 285, No. 47. pp. 36322-36329.

Bibtex - Download

@article{5ff4451c40ea4953990cc3fc57e93b1d,
title = "Roles for Stress-inducible Lambda Glutathione Transferases in Flavonoid Metabolism in Plants as Identified by Ligand Fishing",
abstract = "The glutathione transferases (GSTs) of plants are a superfamily of abundant enzymes whose roles in endogenous metabolism are largely unknown. For example, the lambda class of GSTs (GSTLs) have members that are selectively induced by chemical stress treatments and based on their enzyme chemistry are predicted to have roles in redox homeostasis. However, using conventional approaches these functions have yet to be determined. To address this, recombinant GSTLs from wheat and Arabidopsis were tagged with a Strep tag and after affinity-immobilization, incubated with extracts from Arabidopsis, tobacco, and wheat. Bound ligands were then recovered by solvent extraction and identified by mass spectrometry (MS). With the wheat enzyme TaGSTL1, the ligand profiles obtained with in vitro extracts from tobacco closely matched those observed after the protein had been expressed in planta, demonstrating that these associations were physiologically representative. The stress-inducible TaGSTL1 was found to selectively recognize flavonols (e. g. taxifolin; K-d = 25 nM), with this binding being dependent upon S-glutathionylation of an active site cysteine. In the case of the wheat extracts, this selectivity in ligand recognitions lead to the detection of flavonols that had not been previously described in this cereal. Subsequent in vitro assays showed that the co-binding of flavonols, such as quercetin, to the thiolated TaGSTL1 represented an intermediate step in the reduction of the respective S-glutathionylated quinone derivatives to yield free flavonols. These results suggest a novel role for GSTLs in maintaining the flavonoid pool under stress conditions.",
keywords = "S-TRANSFERASES, ARABIDOPSIS, WHEAT, REGIOSELECTIVITY, REVERSIBILITY, SUPERFAMILY, CONJUGATION, BINDING, FAMILY",
author = "Dixon, {David P.} and Robert Edwards",
year = "2010",
month = "11",
day = "19",
doi = "10.1074/jbc.M110.164806",
language = "English",
volume = "285",
pages = "36322--36329",
journal = "The Journal of biological chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "47",

}

RIS (suitable for import to EndNote) - Download

TY - JOUR

T1 - Roles for Stress-inducible Lambda Glutathione Transferases in Flavonoid Metabolism in Plants as Identified by Ligand Fishing

AU - Dixon, David P.

AU - Edwards, Robert

PY - 2010/11/19

Y1 - 2010/11/19

N2 - The glutathione transferases (GSTs) of plants are a superfamily of abundant enzymes whose roles in endogenous metabolism are largely unknown. For example, the lambda class of GSTs (GSTLs) have members that are selectively induced by chemical stress treatments and based on their enzyme chemistry are predicted to have roles in redox homeostasis. However, using conventional approaches these functions have yet to be determined. To address this, recombinant GSTLs from wheat and Arabidopsis were tagged with a Strep tag and after affinity-immobilization, incubated with extracts from Arabidopsis, tobacco, and wheat. Bound ligands were then recovered by solvent extraction and identified by mass spectrometry (MS). With the wheat enzyme TaGSTL1, the ligand profiles obtained with in vitro extracts from tobacco closely matched those observed after the protein had been expressed in planta, demonstrating that these associations were physiologically representative. The stress-inducible TaGSTL1 was found to selectively recognize flavonols (e. g. taxifolin; K-d = 25 nM), with this binding being dependent upon S-glutathionylation of an active site cysteine. In the case of the wheat extracts, this selectivity in ligand recognitions lead to the detection of flavonols that had not been previously described in this cereal. Subsequent in vitro assays showed that the co-binding of flavonols, such as quercetin, to the thiolated TaGSTL1 represented an intermediate step in the reduction of the respective S-glutathionylated quinone derivatives to yield free flavonols. These results suggest a novel role for GSTLs in maintaining the flavonoid pool under stress conditions.

AB - The glutathione transferases (GSTs) of plants are a superfamily of abundant enzymes whose roles in endogenous metabolism are largely unknown. For example, the lambda class of GSTs (GSTLs) have members that are selectively induced by chemical stress treatments and based on their enzyme chemistry are predicted to have roles in redox homeostasis. However, using conventional approaches these functions have yet to be determined. To address this, recombinant GSTLs from wheat and Arabidopsis were tagged with a Strep tag and after affinity-immobilization, incubated with extracts from Arabidopsis, tobacco, and wheat. Bound ligands were then recovered by solvent extraction and identified by mass spectrometry (MS). With the wheat enzyme TaGSTL1, the ligand profiles obtained with in vitro extracts from tobacco closely matched those observed after the protein had been expressed in planta, demonstrating that these associations were physiologically representative. The stress-inducible TaGSTL1 was found to selectively recognize flavonols (e. g. taxifolin; K-d = 25 nM), with this binding being dependent upon S-glutathionylation of an active site cysteine. In the case of the wheat extracts, this selectivity in ligand recognitions lead to the detection of flavonols that had not been previously described in this cereal. Subsequent in vitro assays showed that the co-binding of flavonols, such as quercetin, to the thiolated TaGSTL1 represented an intermediate step in the reduction of the respective S-glutathionylated quinone derivatives to yield free flavonols. These results suggest a novel role for GSTLs in maintaining the flavonoid pool under stress conditions.

KW - S-TRANSFERASES

KW - ARABIDOPSIS

KW - WHEAT

KW - REGIOSELECTIVITY

KW - REVERSIBILITY

KW - SUPERFAMILY

KW - CONJUGATION

KW - BINDING

KW - FAMILY

UR - http://www.scopus.com/inward/record.url?scp=78449231466&partnerID=8YFLogxK

U2 - 10.1074/jbc.M110.164806

DO - 10.1074/jbc.M110.164806

M3 - Article

VL - 285

SP - 36322

EP - 36329

JO - The Journal of biological chemistry

JF - The Journal of biological chemistry

SN - 0021-9258

IS - 47

ER -