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Year 2013, Volume: 1 Issue: 2, 109 - 117, 01.12.2013

İlknur Özgençli Yusuf Temel Mehmet Çiftçi , Ömer Küfrevioğlu

Abstract

In this study, we determined the in vitro effects of some herbicides and insecticides on the activity of mitochondrial Thioredoxin Reductase that purified from liver of rainbow trout. For purification of enzyme, 2’, 5’-ADP Sepharose 4B affinity chromatography technique was used. The enzyme was purified 648,4-fold from rainbow trout liver mitochondria in a yield of 2.38 % with 11,8 U/mg. SDS-PAGE was done to control the purify of enzyme and also showed a single band for the enzyme. According to the kinetic studies, inhibition effects of all pesticides including four of herbicides and four of insecticides were identified

Keywords

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References

  • Williams, C.H. Jr, Lipoamide dehydrogenase, glutathione reductase, thioredoxinreductase, and mercuric ion reductase, a family of flavoenzyme transhydrogenases, In Chemistry and Biochemistry of Flavoenzymes(Müller, F., ed.), 3; 121-211. CRC Press, Boca Raton, FL., 1992.
  • Powis, G., Montfort, W.R., Properties and biological activities of thioredoxin, Pharmacol Toxicol, 41, 261-295, 2001.
  • Mustacich, D., Powis, G. Thioredoxin reductase, Biochem J, 346, 1-8, 2000.
  • Gallegos, A., Berggren, M.I., Gasdaska, J.R. and Powis, G., Mechanisms of the regulation of thioredoxin reductase activity in cancer cells by the chemopreventive agent selenium, Cancer Res., 57, 4965-4970, 1997.
  • Ayene, IS., Stamato, T.D., Mauldin SK, Biaglow JE, Tuttle SW, Jenkins SF et al., Mutation in the glucose-6-phosphate dehydrogenase gene leads to ınactivation of kudna binding during oxidative stress, J Biol Chem 277, 9929-35, 2002.
  • Turanov, A., Hatfield, D.L.,Gladyshev, V.N., Characterization of Protein Targets of Mammalian Thioredoxin Reductase, Methods enzymol, 474, 245-254, 2010.
  • Nishinaka, Y., Nakamura, H., Masutani, H., Redox control of cellular function by thioredoxin: a new therapeutic direction in host defence, Arch Immunol Ther Exp 49, 285-92, 2001.
  • Arner, E.S.J., Nordberg, J. and Holmgren, A., A. Efficient reduction of lipoamide and lipoic acid by mammalian thioredoxin reductase, Biochem. Biophys. Res. Commun., 225, 268-274, 1996.
  • Björnstedt, M., Hamberg, M., Kumar, S., Xue, J. and Holmgren, A., Human thioredoxin reduces lipid hydroperoxides by NADPH and selenocysteine strongly stimulates the reaction via catalytically generated selenols, J. Biol. Chem., 270, 11761- 11764, 1995.
  • Arner, E.S.J.and Holmgren, A., Physiological functions of thioredoxin and thioredoxin reductase, Eur. J. Biochem., 267, 6102-6109, 2000.
  • Holmgren, A., Reduction of dislufides by thioredoxin. Exceptional reactivity of insulin and suggested functions of thioredoxin in mechanism
  • of hormone action, Eur. J. Biochem., 254, 9113-9119, 1979.
  • Rigobello, M.P., Callegaro, M.T., Barzon, E., Benetti, M.and Bindoli, A., Purification of mitochondrial thioredoxin reductase and ıts ınvolvement ın the regulation of membran permeability, Free Radical Biology- Medicine, 24, 370-376, 1998.
  • May, J.M., Mendiratta, S., Hill, K.E.and Burk, R.F., Reduction of dehydroascorbate to ascorbate by the selenoenzyme thioredoxin reductase, J Biol Chem, 272, 22607- 22610, 1997.
  • May, J.M., Cobb, C.E., Mendiratta, S., Hill, K.E.and Burk, R.F., Reduction of the ascorbyl free radical to ascorbate by thioredoxin reductase, J. Biol. Chem., 273, 23039-23045, 1998.
  • Jordan, A., Reichard, P., Ribonucleotide reductases, Annu Rev Biochem, 67, 71- 98, 1998.
  • Lehninger, A.L., Principles of biochemistry. Newyork: Worth , Publishers Inc, 2000. 117 Özgençli et al. Muş Alparslan University Journal of Science, 1 (2), 109-117, 2013.
  • Bradford, M.M. A rapid and sensitive method for the quantitation(quantifi cation**) of microgram quantities of protein utilizing the principle of protein-dye binding, Anal Biochem, 72, 248-251, 1976.
  • Laemmli, UK., Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-685, 1970.
  • Baker, A., Payne, C.M., Briehi, M.M. and Powis, G.,Thioredoxin, a gene found overexpressed in human cancer, inhibits apoptosis in vitro and in vivo, Cancer Res., 57, 5162-5167, 1997.
  • Gasdaska, J.R., Berggren, M.I., and Powis, G., Cell growth stimulation by the redox protein thioredoxin occurs by a novel helper mechanism, Cell Growth Differ, 6, 1643-1650, 1995.
  • Dündar, Y., Aslan, R., Antioksidan denge ve korunmasında vitaminlerin rolü, Hayvancılık Araştırma Dergisi, 306, 1-17, 1999.
  • Hollstein, M., Sidransky, D., Vogelstein, B., Harris, C.C., P53 mutations in human cancers, Science (Washington DC), 253, 49-53, 1991.
  • Jeorger, A.C., Fersht, A.R., Structure-function- rescue: the diverse nature of common p53 cancer mutants, Oncogene, 26, 2226- 2242, 2007.
  • Hainaut, P., Miller, J., Redox modulation of p53 conformation and sequence-specific DNA binding, Cancer Res., 53, 4469-4473, 1993.
  • Polyak, K., Xia, Y., Zweler, J.L., Kinzler, K.W. and Vogelstein, B., A model for p53-induced apoptosis, Nature (London), 389, 300-303, 1997
  • Kemerdere, R.,Glial tümörlerde tiyoredoksin redüktaz dengeleri (Uzmanlık Tezi),İstanbul Üniversitesi Cerrah Paşa Tıp Fakültesi Nöroşirürji ABD, İstanbul,2008
  • Luthman, M., Holmgren, A., Rat liver thioredoxin and thioredoxin reductase: purification and characterization, Biochemistry, 21, 6628-6633, 1982.
  • Holmgren, A., Bovine thioredoxin system. Purification of thioredoxin reductase from calf liver and thymus and studies of its function in disulfide reduction, J Biol., Chem. 252, 4600-4606, 1977.
  • Gonzales, P.P., Baldesten, A., Reichard, P., Purification of a thioredoxin system from yeast, Journal of Biochem., 245, 2363- 2370, 1970.
  • Bar-Noy, S., Gorlatov, N., Stadtman, T. C.,Overexpression of wild type and secys/cys mutant of human trxr in E.coli: the role of selenocysteine in the catalytic activity,Free Radical Biology&Medicine, 30, 51-61, 2001.
  • Wittle, A.B., Anestal, K., Jerremalm, E., Ehrsson, H., Arner, E.S.J., Inhibition of trxr but not of gr by the major classes of alkylating and platinium-containing anticancer compounds, Free Radical Biology&Medicine, 39, 696-703, 2005.
  • Tandogan, B. and Ulusu,N.N., Thioredoxin reductase, Hacettepe J. Biol, Chem., 39, 87-92, 2011.
  • Carvalho, C. Et. Al, Biomarkers of adverse response to mercury : histopathology versus thioredoxin reductase activity, Journal of Biomedicine and Biotechnology, 359879-359888, 2012

BAZI PESTİSİTLERİN GÖKKUŞAĞI ALABALIĞI KARACİĞERİNDEN SAFLAŞTIRILAN MİTOKONDRİAL TİYOREDOKSİN REDÜKTAZ ENZİMİNİN AKTİVİTESİ ÜZERİNE İN VİTRO ETKİLERİNİN İNCELENMESİ

Year 2013, Volume: 1 Issue: 2, 109 - 117, 01.12.2013

İlknur Özgençli Yusuf Temel Mehmet Çiftçi , Ömer Küfrevioğlu

Abstract

Bu çalışmada, gökkuşağı alabalığı karaciğerinden saflaştırılan mitokondrial tiyoredoksin redüktaz enzimi aktivitesi üzerine bazı herbisit ve insektisitlerin in vitro etkileri incelendi. Enzimin saflaştırılmasında 2’, 5’­ADP Sepharose 4B afinite kromatografisi yöntemi kullanıldı. Enzim 11,8 EÜ/mg protein spesifik aktiviteyle, % 2,38 verimle ve 648,4 kat saflaştırıldı. Enzimin saflık kontrolü Sodyum Dodesil Sülfat-Poliakrilamid Jel Elektroforezi (SDS-PAGE) ile yapıldı ve tüm kinetik çalışmalarda saf enzim kullanıldı. Yapılan kinetik çalış­malarda 4’ü herbisit 4’ü insektisit olmak üzere kullanılan tüm pestisitlerin enzim üzerinde inhibisyon etkisi tespit edildi.

Keywords

Gökkuşağı Alabalığı Tiyoredoksin Redüktaz Herbisit İnsektisit İnhibisyon

References

  • Williams, C.H. Jr, Lipoamide dehydrogenase, glutathione reductase, thioredoxinreductase, and mercuric ion reductase, a family of flavoenzyme transhydrogenases, In Chemistry and Biochemistry of Flavoenzymes(Müller, F., ed.), 3; 121-211. CRC Press, Boca Raton, FL., 1992.
  • Powis, G., Montfort, W.R., Properties and biological activities of thioredoxin, Pharmacol Toxicol, 41, 261-295, 2001.
  • Mustacich, D., Powis, G. Thioredoxin reductase, Biochem J, 346, 1-8, 2000.
  • Gallegos, A., Berggren, M.I., Gasdaska, J.R. and Powis, G., Mechanisms of the regulation of thioredoxin reductase activity in cancer cells by the chemopreventive agent selenium, Cancer Res., 57, 4965-4970, 1997.
  • Ayene, IS., Stamato, T.D., Mauldin SK, Biaglow JE, Tuttle SW, Jenkins SF et al., Mutation in the glucose-6-phosphate dehydrogenase gene leads to ınactivation of kudna binding during oxidative stress, J Biol Chem 277, 9929-35, 2002.
  • Turanov, A., Hatfield, D.L.,Gladyshev, V.N., Characterization of Protein Targets of Mammalian Thioredoxin Reductase, Methods enzymol, 474, 245-254, 2010.
  • Nishinaka, Y., Nakamura, H., Masutani, H., Redox control of cellular function by thioredoxin: a new therapeutic direction in host defence, Arch Immunol Ther Exp 49, 285-92, 2001.
  • Arner, E.S.J., Nordberg, J. and Holmgren, A., A. Efficient reduction of lipoamide and lipoic acid by mammalian thioredoxin reductase, Biochem. Biophys. Res. Commun., 225, 268-274, 1996.
  • Björnstedt, M., Hamberg, M., Kumar, S., Xue, J. and Holmgren, A., Human thioredoxin reduces lipid hydroperoxides by NADPH and selenocysteine strongly stimulates the reaction via catalytically generated selenols, J. Biol. Chem., 270, 11761- 11764, 1995.
  • Arner, E.S.J.and Holmgren, A., Physiological functions of thioredoxin and thioredoxin reductase, Eur. J. Biochem., 267, 6102-6109, 2000.
  • Holmgren, A., Reduction of dislufides by thioredoxin. Exceptional reactivity of insulin and suggested functions of thioredoxin in mechanism
  • of hormone action, Eur. J. Biochem., 254, 9113-9119, 1979.
  • Rigobello, M.P., Callegaro, M.T., Barzon, E., Benetti, M.and Bindoli, A., Purification of mitochondrial thioredoxin reductase and ıts ınvolvement ın the regulation of membran permeability, Free Radical Biology- Medicine, 24, 370-376, 1998.
  • May, J.M., Mendiratta, S., Hill, K.E.and Burk, R.F., Reduction of dehydroascorbate to ascorbate by the selenoenzyme thioredoxin reductase, J Biol Chem, 272, 22607- 22610, 1997.
  • May, J.M., Cobb, C.E., Mendiratta, S., Hill, K.E.and Burk, R.F., Reduction of the ascorbyl free radical to ascorbate by thioredoxin reductase, J. Biol. Chem., 273, 23039-23045, 1998.
  • Jordan, A., Reichard, P., Ribonucleotide reductases, Annu Rev Biochem, 67, 71- 98, 1998.
  • Lehninger, A.L., Principles of biochemistry. Newyork: Worth , Publishers Inc, 2000. 117 Özgençli et al. Muş Alparslan University Journal of Science, 1 (2), 109-117, 2013.
  • Bradford, M.M. A rapid and sensitive method for the quantitation(quantifi cation**) of microgram quantities of protein utilizing the principle of protein-dye binding, Anal Biochem, 72, 248-251, 1976.
  • Laemmli, UK., Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-685, 1970.
  • Baker, A., Payne, C.M., Briehi, M.M. and Powis, G.,Thioredoxin, a gene found overexpressed in human cancer, inhibits apoptosis in vitro and in vivo, Cancer Res., 57, 5162-5167, 1997.
  • Gasdaska, J.R., Berggren, M.I., and Powis, G., Cell growth stimulation by the redox protein thioredoxin occurs by a novel helper mechanism, Cell Growth Differ, 6, 1643-1650, 1995.
  • Dündar, Y., Aslan, R., Antioksidan denge ve korunmasında vitaminlerin rolü, Hayvancılık Araştırma Dergisi, 306, 1-17, 1999.
  • Hollstein, M., Sidransky, D., Vogelstein, B., Harris, C.C., P53 mutations in human cancers, Science (Washington DC), 253, 49-53, 1991.
  • Jeorger, A.C., Fersht, A.R., Structure-function- rescue: the diverse nature of common p53 cancer mutants, Oncogene, 26, 2226- 2242, 2007.
  • Hainaut, P., Miller, J., Redox modulation of p53 conformation and sequence-specific DNA binding, Cancer Res., 53, 4469-4473, 1993.
  • Polyak, K., Xia, Y., Zweler, J.L., Kinzler, K.W. and Vogelstein, B., A model for p53-induced apoptosis, Nature (London), 389, 300-303, 1997
  • Kemerdere, R.,Glial tümörlerde tiyoredoksin redüktaz dengeleri (Uzmanlık Tezi),İstanbul Üniversitesi Cerrah Paşa Tıp Fakültesi Nöroşirürji ABD, İstanbul,2008
  • Luthman, M., Holmgren, A., Rat liver thioredoxin and thioredoxin reductase: purification and characterization, Biochemistry, 21, 6628-6633, 1982.
  • Holmgren, A., Bovine thioredoxin system. Purification of thioredoxin reductase from calf liver and thymus and studies of its function in disulfide reduction, J Biol., Chem. 252, 4600-4606, 1977.
  • Gonzales, P.P., Baldesten, A., Reichard, P., Purification of a thioredoxin system from yeast, Journal of Biochem., 245, 2363- 2370, 1970.
  • Bar-Noy, S., Gorlatov, N., Stadtman, T. C.,Overexpression of wild type and secys/cys mutant of human trxr in E.coli: the role of selenocysteine in the catalytic activity,Free Radical Biology&Medicine, 30, 51-61, 2001.
  • Wittle, A.B., Anestal, K., Jerremalm, E., Ehrsson, H., Arner, E.S.J., Inhibition of trxr but not of gr by the major classes of alkylating and platinium-containing anticancer compounds, Free Radical Biology&Medicine, 39, 696-703, 2005.
  • Tandogan, B. and Ulusu,N.N., Thioredoxin reductase, Hacettepe J. Biol, Chem., 39, 87-92, 2011.
  • Carvalho, C. Et. Al, Biomarkers of adverse response to mercury : histopathology versus thioredoxin reductase activity, Journal of Biomedicine and Biotechnology, 359879-359888, 2012
There are 34 citations in total.

Details

Primary Language Turkish
Journal Section Research Article
Authors

İlknur Özgençli This is me

Yusuf Temel This is me

Mehmet Çiftçi

Ömer Küfrevioğlu This is me

Publication Date December 1, 2013
Published in Issue Year 2013 Volume: 1 Issue: 2

Cite

APA Özgençli, İ., Temel, Y., Çiftçi, M., Küfrevioğlu, Ö. (2013). BAZI PESTİSİTLERİN GÖKKUŞAĞI ALABALIĞI KARACİĞERİNDEN SAFLAŞTIRILAN MİTOKONDRİAL TİYOREDOKSİN REDÜKTAZ ENZİMİNİN AKTİVİTESİ ÜZERİNE İN VİTRO ETKİLERİNİN İNCELENMESİ. Mus Alparslan University Journal of Science, 1(2), 109-117. https://doi.org/10.18586/msufbd.92286
AMA Özgençli İ, Temel Y, Çiftçi M, Küfrevioğlu Ö. BAZI PESTİSİTLERİN GÖKKUŞAĞI ALABALIĞI KARACİĞERİNDEN SAFLAŞTIRILAN MİTOKONDRİAL TİYOREDOKSİN REDÜKTAZ ENZİMİNİN AKTİVİTESİ ÜZERİNE İN VİTRO ETKİLERİNİN İNCELENMESİ. MAUN Fen Bil. Dergi. December 2013;1(2):109-117. doi:10.18586/msufbd.92286
Chicago Özgençli, İlknur, Yusuf Temel, Mehmet Çiftçi, and Ömer Küfrevioğlu. “BAZI PESTİSİTLERİN GÖKKUŞAĞI ALABALIĞI KARACİĞERİNDEN SAFLAŞTIRILAN MİTOKONDRİAL TİYOREDOKSİN REDÜKTAZ ENZİMİNİN AKTİVİTESİ ÜZERİNE İN VİTRO ETKİLERİNİN İNCELENMESİ”. Mus Alparslan University Journal of Science 1, no. 2 (December 2013): 109-17. https://doi.org/10.18586/msufbd.92286.
EndNote Özgençli İ, Temel Y, Çiftçi M, Küfrevioğlu Ö (December 1, 2013) BAZI PESTİSİTLERİN GÖKKUŞAĞI ALABALIĞI KARACİĞERİNDEN SAFLAŞTIRILAN MİTOKONDRİAL TİYOREDOKSİN REDÜKTAZ ENZİMİNİN AKTİVİTESİ ÜZERİNE İN VİTRO ETKİLERİNİN İNCELENMESİ. Mus Alparslan University Journal of Science 1 2 109–117.
IEEE İ. Özgençli, Y. Temel, M. Çiftçi, and Ö. Küfrevioğlu, “BAZI PESTİSİTLERİN GÖKKUŞAĞI ALABALIĞI KARACİĞERİNDEN SAFLAŞTIRILAN MİTOKONDRİAL TİYOREDOKSİN REDÜKTAZ ENZİMİNİN AKTİVİTESİ ÜZERİNE İN VİTRO ETKİLERİNİN İNCELENMESİ”, MAUN Fen Bil. Dergi., vol. 1, no. 2, pp. 109–117, 2013, doi: 10.18586/msufbd.92286.
ISNAD Özgençli, İlknur et al. “BAZI PESTİSİTLERİN GÖKKUŞAĞI ALABALIĞI KARACİĞERİNDEN SAFLAŞTIRILAN MİTOKONDRİAL TİYOREDOKSİN REDÜKTAZ ENZİMİNİN AKTİVİTESİ ÜZERİNE İN VİTRO ETKİLERİNİN İNCELENMESİ”. Mus Alparslan University Journal of Science 1/2 (December 2013), 109-117. https://doi.org/10.18586/msufbd.92286.
JAMA Özgençli İ, Temel Y, Çiftçi M, Küfrevioğlu Ö. BAZI PESTİSİTLERİN GÖKKUŞAĞI ALABALIĞI KARACİĞERİNDEN SAFLAŞTIRILAN MİTOKONDRİAL TİYOREDOKSİN REDÜKTAZ ENZİMİNİN AKTİVİTESİ ÜZERİNE İN VİTRO ETKİLERİNİN İNCELENMESİ. MAUN Fen Bil. Dergi. 2013;1:109–117.
MLA Özgençli, İlknur et al. “BAZI PESTİSİTLERİN GÖKKUŞAĞI ALABALIĞI KARACİĞERİNDEN SAFLAŞTIRILAN MİTOKONDRİAL TİYOREDOKSİN REDÜKTAZ ENZİMİNİN AKTİVİTESİ ÜZERİNE İN VİTRO ETKİLERİNİN İNCELENMESİ”. Mus Alparslan University Journal of Science, vol. 1, no. 2, 2013, pp. 109-17, doi:10.18586/msufbd.92286.
Vancouver Özgençli İ, Temel Y, Çiftçi M, Küfrevioğlu Ö. BAZI PESTİSİTLERİN GÖKKUŞAĞI ALABALIĞI KARACİĞERİNDEN SAFLAŞTIRILAN MİTOKONDRİAL TİYOREDOKSİN REDÜKTAZ ENZİMİNİN AKTİVİTESİ ÜZERİNE İN VİTRO ETKİLERİNİN İNCELENMESİ. MAUN Fen Bil. Dergi. 2013;1(2):109-17.