A Glutathione S-transferase Elutes with Glyoxalase-I (Gly-I) During Purification of Gly-I from Maize (Zea mays L.) Seedlings
Journal of Plant Sciences
Volume 4, Issue 1, February 2016, Pages: 8-12
Received: Dec. 27, 2015;
Accepted: Jan. 8, 2016;
Published: Jan. 21, 2016
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Md. Motiar Rohman, Molecular Breeding Lab, Plant Breeding Division, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh
Afsana Hoque Akhi, Molecular Breeding Lab, Plant Breeding Division, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh
Nusrat Jahan Methela, Department of Genetics and Plant Breeding, Patuakhali Science and Technology University, Patuakhali, Bangladesh
Mohammad Golam Hossain, Molecular Breeding Lab, Plant Breeding Division, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh
M. Shalim Uddin, Molecular Breeding Lab, Plant Breeding Division, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh
Mohammad Amiruzzaman, Molecular Breeding Lab, Plant Breeding Division, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh
Bhagya Rani Banik, Training and Communication Wing, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh
In this study an attempt was taken to purify Glyoxalase-I (Gly-I: E.C., 188.8.131.52), from maize seedlings. Both green and non-green parts of 7 day old maize seedlings were used as plant materials. Crude proteins were precipitated by 65% (NH4)2SO4, and dialyzed overnight. The dialyzate was applied on DEAE-cellulose chromatography and eluted with linear gradient of KCl from 0 to 0.2 M. In both cases, Gly-I eluted at approximately 85 mM of KCL. The active Gly-I fractions were pooled and applied on a hydroxylapatite chromatography and eluted with 0-40 mM potassium-phosphate buffer, but the eluted fractions showed very poor activity. Therefore, the active pooled fraction of DEAE-chromatography was then applied directly on affinity chromatography (S-hexyl glutathione-agarose) for final purification and eluted with 1.2 mM of S-hexyl glutathione. The purified protein from green and non-green part had specific activity of 33.23 and 39.25 μmol min-1 mg-1 protein, respectively, along with recovery of 1.47 and 162, respectively, and yield of 83.11 and 68.15, respectively. In SDS-PAGE, the active purified affinity fraction was found to move with another protein. The spectrophotometric analysis of high active Gly-I fractions from DEAE-cellulose and affinity chromatography showed GST [another detoxifying enzyme (E.C., 184.108.40.206)] activity. This result suggested that one of the adjacent protein bands in SDS-PAGE was due to presence of a GST in Gly-I fraction.
Md. Motiar Rohman,
Afsana Hoque Akhi,
Nusrat Jahan Methela,
Mohammad Golam Hossain,
M. Shalim Uddin,
Bhagya Rani Banik,
A Glutathione S-transferase Elutes with Glyoxalase-I (Gly-I) During Purification of Gly-I from Maize (Zea mays L.) Seedlings, Journal of Plant Sciences.
Vol. 4, No. 1,
2016, pp. 8-12.
Wang W, Vinocur B, Altman A. 2003. Plant responses to drought, salinity and extreme temperatures towards genetic engineering for stress tolerance. Planta 218: 1-14.
Gill SS, Tuteja N. 2010. Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol. Biochem 48: 909-930.
Ahuja I, de Vos RCH, Bones AM. Hall RD. 2010. Plant molecular stress responses face climate change. Trends in Plant Sci 15(12): 664-674.
Veena, Reddy VS, Sopory SK. 1999. Glyoxalase I from Brassica juncea: molecular cloning, regulation and its over-expression confer tolerance in transgenic tobacco under stress. The Plant Journal 17: 385-395.
Chen ZY, Brown RL, Damann KE, Cleveland TE. 2004. Identification of a maize kernel stress-related protein and its effect on aflatoxin accumulation. Phytopathology 94: 938-945.
Yadav SK, Singla-Pareek SL, Ray M, Reddy MK, Sopory SK. 2005. Methylglyoxal levels in plants under salinity stress are dependent on glyoxalase I and glutathione. Biochem. Biophysic. Res. Commun 337: 61-67.
Yadav SK, Singla-Pareek SL, Ray M, Reddy MK, Sopory SK. 2005. Transgenic tobacco plants overexpressing glyoxalase enzymes resist an increase in methylglyoxal and maintain higher reduced glutathione levels under salinity stress. FEBS Letters 579: 6265-6271.
Singla-Pareek SL, Yadav SK, Pareek A, Reddy MK, Sopory SK. 2006. Transgenic tobacco overexpressing glyoxalase pathway enzymes grow and set viable seeds in zinc-spiked soils. Plant Physiol 140(2): 613-623.
Banu MNA, Hoque MA, Watamable-Sugimoto M, Islam MA, Uraji M, Matsuoka M, Nakamura Y, Murata Y. 2010. Proline and glycinebetaine ameliorated NaCl stress via scavenging of hydrogen peroxide and methylglyoxal but not superoxide or nitric oxide in tobacco cultured cells. Biosci. Biotechnol. Biochem 74: 2043-2049.
El-Shabrawi H, Kumar B, Kaul T, Reddy MK, Singla-Pareek SL, Sopory, SK. 2010. Redox homeostasis, antioxidant defense, and methylglyoxal detoxification as markers for salt tolerance in Pokkali rice. Protoplasma. 245: 85-96.
Noctor G, Mhamdi A, Chaouch S, Han Y, Neukermans J,Marquez-Garcia B, Queval G,Foyer CH. 2012. Glutathione in plants: an integrated overview. Plant Cell Environ 35: 454-484.
Rohman MM, Molla MR, Rahman MM Ahmed A, Biswas A, Amiruzzaman M. 2015. Proline and betaine upregulated glutathione dependent detoxification enzymes in tolerant maize seedlings under saline stress. J. Plant Sci 3(6): 294-302.
Chakravarty TN, Sopory SK. 1998. Blue light stimulation of cell proliferation and glyoxalase I activity in callus cultures of Amaranthus paniculatus. Plant Sci 132: 63-69.
Deswal R, Chakravarty TN, Sopory SK. 1993. The glyoxalase system in higher plants: regulation in growth and differentiation. Biochem. Soc. Trans 21:527-530.
Paulus C, Kollner B, Jacobsen HJ. 1993. Physiological and biochemical characterization of glyoxalase I: a general marker for cell proliferation from a soybean cell suspension. Planta 189: 561-566.
Sethi U, Basu A and Guha-Mukherjee S. 1988. Control of cell proliferation and differentiation by regulating polyamine biosynthesis in cultures of Brassica and its correlation with glyoxalase I activity. Plant Sci 56:167-175.
Espartero J, Aguayo IS, Pardo JM. 1995. Molecular characterization of glyoxalase-I from a higher plant; upregulation by stress. Plant Molecular Biology 29: 1223-1233.
Singla-Pareek SL, Ray M, Reddy MK, Sopory SK. 2003. Genetic engineering of the glyoxalase pathway in tobacco leads to enhanced salinity tolerance. Proceedings of the Natl. Acad.Sci USA 100: 14672-14677.
Saxena M, Bisht R, Roy DS, Sopory SK, Bhalla-Sarinn M. 2005. Cloning and characterization of a mitochondrial glyoxalase II from Brassica juncea that is upregulated by NaCl, Zn and ABA. Biochem. Biophysic. Res. Commun 336: 813-819.
Hoque MA, Banu MNA, Nakamura Y, Shimoishi Y, Murata Y. 2008. Proline and glycinebetaine enhance antioxidant defense and methylglyoxal detoxification systems and reduce NaCl-induced damage in cultured tobacco cells. J. Plant Physiol. 165: 813-824.
Hoque MA, Okuma E, Banu MNA, Nakamura Y, Shimoishi Y, Murata Y. 2007. Exogenous proline mitigates the detrimental effects of salt stress more than exogenous betaine by increasing antioxidant enzyme activities. J. Plant Physiol 164 (5): 553-61.
Hoque MA, Banu MNA, Okuma E, Amako K, Nakamura Y, Shimoishi Y, Murata Y. 2007. Exogenous proline and glycinebetaine ingresses NaCl-induced ascorbate-glutathione cycle enzyme activities and proline improves salt tolerance more than glycinebetaine in tobacco Bright yellow-2 suspension-cultured cells. J. Plant Physiol 164 (11): 1457-1468.
Bradford MM. 1976. A rapid and sensitive for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem72: 248-54.
Laemmli UK. 1970. Cleavage of structural proteins during the assembly of head of bacteriophage T4. Nature 227: 680-685.
Rohman MM, Uddin MS, Fujita M. 2010. Up-regulation of onion bulb glutathione S-transferases (GSTs) by abiotic stresses: A comparative study between two differently sensitive GSTs to their physiological inhibitors. Plant Omics J 3 (1): 28-34.
Rohman MM, Begum S, Akhi AH, Ahsan AFMS, Uddin MS, Amiruzzaman M, Banik BR.2015. Protective role of antioxidants in maize seedlings undersaline stress: Exogenous proline provided better tolerance than betaine. Bothalia J 45(4): 17-35.
Deswal R and Sopory SK. 1999.Glyoxalase I from Brassica juncea is a calmodulin stimulated protein. Biochimica et Biophysica Acta 1450: 460-467.
Islam MR, Chowdhury K, Rahman MM, Rohman MM. 2015. Comparative investigation of glutathione S-transferase (GST) in different crops and purification of high active GSTs from onion (Allium cepa L.). J. Plant Sci 3: 162-170.