|
|
|
|
|
| Expression and Characterization of a Sigma-Class Glutathione S-transferase of the Oriental Migratory Locust, Locusta migratoria manilensis (Meyen) |
| JIA Miao, QIN Guo-hua, LIU Ting, ZHANG Jian-zhen, ZHANG Xue-yao, ZHU Kun-yan, GUO Yaping, MA En-bo |
1.Research Institute of Applied Biology, Shanxi University
2.College of Environmental Science and Resource, Shanxi University
3.Department of Entomology, 123 Waters Hall, Kansas State University
4.School of Life Science, Shanxi University |
|
|
|
摘要 A cDNA encoding a sigma-class glutathione S-transferase of the locust, Locusta migratoria manilensis (LmGSTs1), was cloned by reverse transcriptase-polymerase chain reaction. The 830 bp-long cDNA encoded a 615 bp open reading frame (204 amino acid polypeptide), which exhibited the structural motif and domain organization characteristic of GST sigmaclass. It revealed 59, 57, 57, and 56% identities to sigma-class GSTs from Blattella germanica, Gryllotalpa orientalis, Nasonia vitripennis, and Pediculus humanus corporis, respectively. A recombinant protein (LmGSTs1) was functionally expressed in Escherichia coli cells in a soluble form and purified to homogeneity. LmGSTs1 was able to catalyze the biotranslation of glutathione with 1-chloro-2,4-dinitrobenzene, a model substrate for GSTs, as well as with p-nitro-benzyl chloride. Its optimal activity was observed at pH 8.0 and at 30°C. Incubation for 30 min at temperatures below 50°C scarcely affected the activity. The I50 of reactive blue (RB) was 18.5 μmol L-1. In the presence of 0.05 mmol L-1 ethacrynic acid (ECA), LmGSTs1 showed (81±3)% of the original activities.
Abstract A cDNA encoding a sigma-class glutathione S-transferase of the locust, Locusta migratoria manilensis (LmGSTs1), was cloned by reverse transcriptase-polymerase chain reaction. The 830 bp-long cDNA encoded a 615 bp open reading frame (204 amino acid polypeptide), which exhibited the structural motif and domain organization characteristic of GST sigmaclass. It revealed 59, 57, 57, and 56% identities to sigma-class GSTs from Blattella germanica, Gryllotalpa orientalis, Nasonia vitripennis, and Pediculus humanus corporis, respectively. A recombinant protein (LmGSTs1) was functionally expressed in Escherichia coli cells in a soluble form and purified to homogeneity. LmGSTs1 was able to catalyze the biotranslation of glutathione with 1-chloro-2,4-dinitrobenzene, a model substrate for GSTs, as well as with p-nitro-benzyl chloride. Its optimal activity was observed at pH 8.0 and at 30°C. Incubation for 30 min at temperatures below 50°C scarcely affected the activity. The I50 of reactive blue (RB) was 18.5 μmol L-1. In the presence of 0.05 mmol L-1 ethacrynic acid (ECA), LmGSTs1 showed (81±3)% of the original activities.
|
|
Received: 06 September 2010
Accepted: 18 October 2011
|
| Fund: Our research was supported by the Funds for International Cooperation and Exchange of the National Natural Science Foundation of China (30810103907), the National Natural Science Foundation of China (30870302), the Public Welfare Fund for Agriculture, Ministry of Agriculture, China (200903021), and the China Postdoctoral Science Foundation (Special Program, 201003656; Regular Program, 20090451359). |
|
Corresponding Authors:
Correspondence MA En-bo, Professor, Tel/Fax: +86-351-7018871, E-mail: maenbo2003@sxu.edu.cn
E-mail: maenbo2003@sxu.edu.cn
|
| About author: JIA Miao, MSc candidate, E-mail: jiamiao00013@163.com |
Cite this article:
JIA Miao, QIN Guo-hua, LIU Ting, ZHANG Jian-zhen, ZHANG Xue-yao, ZHU Kun-yan, GUO Yaping, MA En-bo.
2011.
Expression and Characterization of a Sigma-Class Glutathione S-transferase of the Oriental Migratory Locust, Locusta migratoria manilensis (Meyen) . Journal of Integrative Agriculture, 10(10): 1570-1576.
|
| [1]Agianian B, Tucker P A, Schouten A, Leonard K, Bullard B, Gros P. 2003. Structure of a Drosophila sigma class glutathione S-transferase reveals a novel active site topography suited for lipid peroxidation products. Journal of Molecular Biology, 326, 151-165. [2]Allocati N, Federici L, Masulli M, Di Ilio C. 2009. Glutathione transferases in bacteria. FEBS Letters, 276, 58-75. [3]Arruda L K, Vailes L D, PlattS-Mills T A, Hayden M L, Chapman M D. 1997. Induction of IgE antibody responses by glutathione S-transferase from the German cockroach (Blattella germanica). Journal of Biological Chemistry, 272, 20907-20912. [4]Bradford M M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248-254. [5]Chen Y L. 2000. The characteristic, cause and ecological management of locust disaster. Bulletin Biology, 35, 1-5. [6]Habig W H, Pabst M J, Jakoby W B. 1974. Glutathione Stransferases. Journal of Biological Chemistry, 249, 7130- 7139. [7]Kang L, Chen X Y, Zhou Y, Liu B W, Zheng W, Li R Q, Wang J, Yu J. 2004. The analysis of large-scale gene expression correlated to the phase changes of the migratory locust. Proceedings of the National Academy of Sciences of the USA, 101, 17611-17615. [8]Masella R, Benedetto R D, Vari R, Filesi C, Giovannini C. 2005. Novel mechanisms of natural antioxidant compounds in biological systems: involvement of glutathione and glutathonerelated enzymes. Journal of Nutritional Biochemistry, 16, 577-586. [9]Papadopoulos A I, Polemitou I, Laifi P, Yiangou A, Tananaki C. 2004. Glutathione S-transferase in the insect Apis mellifera macedonica kinetic characteristics and effect of stress on the expression of GST isoenzymes in the adult worker bee. Comparative Biochemistry and Physiology (C-Toxicology & Pharmacology), 139, 93-97. [10]Ranson H, Rossiter L, Ortelli F, Jensen B, Wang X, Roth C W, Collins F H, Hemingway J. 2001. Identification of a novel class of insect glutathione S-transferases involved in resistance to DDT in the malaria vector Anopheles gambiae. Biochemical Journal, 359, 295-304. [11]Ranson H, Hemingway J. 2005. Mosquito glutathione transferases. Methods in Enzymology, 401, 226-241. [12]Rauch N, Nauen R. 2004. Characterization and molecular cloning of a glutathione S-transferase from the whitefly Bemisia tabaci (Hemiptera: Aleyrodidae). Insect Biochemistry and Molecular Biology, 34, 321-329. [13]Saitou N, Nei M. 1987. The neighbour-joining method: a new method for reconstructing phylogenetic trees. Molecular Biolology and Evolution, 4, 406-425. [14]Thompson J D, Higgins D G, Gibson T J. 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research, 22, 4673-4680. [15]Valles S M, Perera O P, Strong C A. 2003. Purification, biochemical characterization, and cDNA cloning of a glutathione Stransferase from the red imported fire ant, Solenopsis invicta. Insect Biochemistry and Molecular Biology, 33, 981-988. [16]Vontas J G, Small G J, Nikou D C, Ranson H, Hemingway J. 2002. Purification, molecular cloning and heterologous expression of a glutathione S-transferase involved in insecticide resistance from the rice brown planthopper, Nilaparvata lugens. Biochemical Journal, 362, 329-337. [17]Wan Q, Whang I, Lee J S, Lee J. 2009. Novel omega glutathione S-transferases in disk abalone: Characterization and protective roles against environmental stress. Comparative Biochemistry and Physiology (C-Toxicology & Pharmacology), 150, 558- 568. [18]Yamamoto K, Fujii H, Aso Y, Banno Y, Koga K. 2007. Expression and characterization of a sigma-class glutathione S-transferase of the fall webworm, Hyphantria cunea. Bioscience Biotechnology and Biochemistry, 71, 553-560.[19]Yamamoto K, Zhang P, Miake F, Kashige N, Aso Y, Banno Y, Fujii H. 2005. Cloning, expression and characterization of theta-class glutathione S-transferase from the silkworm, Bombyx mori. Comparative Biochemistry and Physiology (BBiochemistry & Molecular Biology), 141, 340-346. [20]Yang M L, Zhang J Z, Zhu K Y, Xuan T, Liu X J, Guo Y P, Ma E B. 2009. Mechanisms of organophosphate resistance in a field population of oriental migratory locust, Locusta migratoria manilensis (Meyen). Archives of Insect Biochemistry and Physiology, 71, 3-15. [21]Yu Q, Lu C, Li B, Fang S, Zuo W, Dai F, Zhang Z, Xiang Z. 2008. Identification, genomic organization and expression pattern of glutathione S-transferase in the silkworm, Bombyx mori. Insect Biochemistry and Molecular Biology, 38, 1158-1164. [22]Yu S J, Huang S W. 2000. Purification and characterization of glutathione S-transferases from the german cockroach, Blattella germanica (L.). Pesticide Biochemistry and Physiology, 67, 36-45. [23]Yu S J. 2002. Biochemical characteristics of microsomal and cytosolic glutathione S-transferases in larvae of the fall armyworm, Spodoptera frugiperda (J. E. Smith). Pesticide Biochemistry and Physiology, 72, 100-110. |
| No Suggested Reading articles found! |
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
