Purification and characterization of glutathione S-transferase from sugarcane leaves

Sharad S. Singhal; Narendra K. Tiwari; Hassan Ahmad; Satish K. Srivastava; Yogesh C. Awasthi

doi:10.1016/0031-9422(91)84175-R

Purification and characterization of glutathione S-transferase from sugarcane leaves

Sharad S. Singhal, Narendra K. Tiwari, Hassan Ahmad, Satish K. Srivastava, Yogesh C. Awasthi

Research output: Contribution to journal › Article › peer-review

19 Scopus citations

Abstract

Glutathione S-transferase has been purified from leaves of sugarcane using (NH₄)₂SO₄ fractionation, isoelectric focusing, Sephadex G-100 gel filtration, DEAE cellulose ion-exchange chromatography, and affinity chromatography over GSH-linked to epoxy-activated Sepharose 6B. The enzyme has a pI value of 4.7 and consists of two subunits having M_r values of ca 22 500 and 24 000. N-Terminal amino acid sequence analysis of both the subunits indicates a high degree of homology between these two subunits. Both subunits also have primary structure homologies with the corn GST subunits characterized previously. Kinetic properties of the enzyme are described and compared with other plant glutathione S-transferases. Similar to human GST isoenzymes, sugarcane leaf GST is also inhibited by the herbicides 2,4-dichlorophenoxyacetate, 2,4,5-trichlorophenoxyacetate, and chenodeoxycholate.

Original language	English (US)
Pages (from-to)	1409-1414
Number of pages	6
Journal	Phytochemistry
Volume	30
Issue number	5
DOIs	https://doi.org/10.1016/0031-9422(91)84175-R
State	Published - 1991
Externally published	Yes

Keywords

Gramineae
Saacharum officinarum
glutathione
glutathione S-transferase.
sugarcane

ASJC Scopus subject areas

Biochemistry
Molecular Biology
Plant Science
Horticulture

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10.1016/0031-9422(91)84175-R

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@article{ad95cedd8f79441b85775e72b8178385,

title = "Purification and characterization of glutathione S-transferase from sugarcane leaves",

abstract = "Glutathione S-transferase has been purified from leaves of sugarcane using (NH4)2SO4 fractionation, isoelectric focusing, Sephadex G-100 gel filtration, DEAE cellulose ion-exchange chromatography, and affinity chromatography over GSH-linked to epoxy-activated Sepharose 6B. The enzyme has a pI value of 4.7 and consists of two subunits having Mr values of ca 22 500 and 24 000. N-Terminal amino acid sequence analysis of both the subunits indicates a high degree of homology between these two subunits. Both subunits also have primary structure homologies with the corn GST subunits characterized previously. Kinetic properties of the enzyme are described and compared with other plant glutathione S-transferases. Similar to human GST isoenzymes, sugarcane leaf GST is also inhibited by the herbicides 2,4-dichlorophenoxyacetate, 2,4,5-trichlorophenoxyacetate, and chenodeoxycholate.",

keywords = "Gramineae, Saacharum officinarum, glutathione, glutathione S-transferase., sugarcane",

author = "Singhal, {Sharad S.} and Tiwari, {Narendra K.} and Hassan Ahmad and Srivastava, {Satish K.} and Awasthi, {Yogesh C.}",

note = "Funding Information: Acknowledgements-- This work was supported in part by USPHS Grant GM-32304, awarded by the National Institute of Health and EY 04396, awarded by the National Eye Institute. We would like to thank Dr Richard R. Fritz for help in the amino acid sequencing.T he help of Mrs Sonya Womack in preparation of this manuscript is acknowledged.",

year = "1991",

doi = "10.1016/0031-9422(91)84175-R",

language = "English (US)",

volume = "30",

pages = "1409--1414",

journal = "Phytochemistry",

issn = "0031-9422",

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number = "5",

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AU - Singhal, Sharad S.

AU - Tiwari, Narendra K.

AU - Ahmad, Hassan

AU - Srivastava, Satish K.

AU - Awasthi, Yogesh C.

N1 - Funding Information: Acknowledgements-- This work was supported in part by USPHS Grant GM-32304, awarded by the National Institute of Health and EY 04396, awarded by the National Eye Institute. We would like to thank Dr Richard R. Fritz for help in the amino acid sequencing.T he help of Mrs Sonya Womack in preparation of this manuscript is acknowledged.

PY - 1991

Y1 - 1991

N2 - Glutathione S-transferase has been purified from leaves of sugarcane using (NH4)2SO4 fractionation, isoelectric focusing, Sephadex G-100 gel filtration, DEAE cellulose ion-exchange chromatography, and affinity chromatography over GSH-linked to epoxy-activated Sepharose 6B. The enzyme has a pI value of 4.7 and consists of two subunits having Mr values of ca 22 500 and 24 000. N-Terminal amino acid sequence analysis of both the subunits indicates a high degree of homology between these two subunits. Both subunits also have primary structure homologies with the corn GST subunits characterized previously. Kinetic properties of the enzyme are described and compared with other plant glutathione S-transferases. Similar to human GST isoenzymes, sugarcane leaf GST is also inhibited by the herbicides 2,4-dichlorophenoxyacetate, 2,4,5-trichlorophenoxyacetate, and chenodeoxycholate.

AB - Glutathione S-transferase has been purified from leaves of sugarcane using (NH4)2SO4 fractionation, isoelectric focusing, Sephadex G-100 gel filtration, DEAE cellulose ion-exchange chromatography, and affinity chromatography over GSH-linked to epoxy-activated Sepharose 6B. The enzyme has a pI value of 4.7 and consists of two subunits having Mr values of ca 22 500 and 24 000. N-Terminal amino acid sequence analysis of both the subunits indicates a high degree of homology between these two subunits. Both subunits also have primary structure homologies with the corn GST subunits characterized previously. Kinetic properties of the enzyme are described and compared with other plant glutathione S-transferases. Similar to human GST isoenzymes, sugarcane leaf GST is also inhibited by the herbicides 2,4-dichlorophenoxyacetate, 2,4,5-trichlorophenoxyacetate, and chenodeoxycholate.

KW - Gramineae

KW - Saacharum officinarum

KW - glutathione

KW - glutathione S-transferase.

KW - sugarcane

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Purification and characterization of glutathione S-transferase from sugarcane leaves

Abstract

Keywords

ASJC Scopus subject areas

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