Araştırma Makalesi
SAFLAŞTIRILAN MİTOKONDRİAL TİYOREDOKSİN REDÜKTAZ ENZİMİNİN AKTİVİTESİ ÜZERİNE İN VİTRO ETKİLERİNİN İNCELENMESİ
Öz
Bu çalışmada, hindi (Meleagris gallopavö) karaciğerinden
saflaştırılan mitokondriyal tiyoredoksin redüktaz enzimi
aktivitesi üzerine bazı herbisit ve insektisitlerin in vitro etkileri
incelendi. Enzimin saflaştırılmasında gradientli santrifügasyonla
homojenizasyondan sonra 2', 5'-ADP Sepharose 4B afinite kromatografisi yöntemi
kullanıldı. Enzim hindi karaciğer mitokondrisinden 3,07 EÜ/mg protein spesifik aktiviteyle, % 2,05 verimle, ve
990 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
kullanılan 4 herbisit ve 4 insektisitten 3 herbisit ve 3 insektisitin
enzim aktivitesi üzerinde inhibisyon etkisi olduğu tespit edildi.
Anahtar Kelimeler
Kaynakça
- [1]. Arner, E. S. and Holmgren, A., Physiological functions of thioredoxin and thioredoxin reductase, Eur. J. Biochem., 267, 6102-6109, 2000. [2]. Söderberg, A., Sahaf, B., Rosen, A., Thioredoxin reductase, a redox active selenoprotein, is secreted by normal and neoplastic cells: presence in human plasma, Cancer research, 60, 2281-2289, 2000. [3]. Lincoln, D.T., Emadi, E.M., Tonissen, K.F., Clarke FM. Thethioredoxin-thioredoxin reductase system: over-expression inhuman cancer, Anticancer Res., 23(3b), 2425-33,2003. [4]. Williams Jr, C. H. Lipoamide dehydrogenase, glutathione reductase, thioredoxin reductase, and mercuric ion reductase a family of flavoenzyme transhydrogenases. Chemistry and biochemistry of flavoenzymes, 3, 121-211, 1992.
- [5]. Powis, G., Montfort, W.R, Properties and biological activities of thioredoxin, Pharmacol Toxicol, 41, 261 -295, 2001. [6]. Mustacich, D., Powis, G., Thioredoxin reductese. BiochemJ. 346,1-8,2000. [7]. Gallegos, A., Berggren, M.I., Gasdaska, J.R. and Powis, G., Mechanisms of the regulation of thioredoxin reductase activity in cancer cells by the chemo preventive agent selenium, Cancer Res., 57,4965-4970,1997. [8]. Ayene, I.S., Stamato, T.D., Mauldin, S.K., Biaglow, J.E., Tuttle, S.W., Jenkins, S.F, Mutation in the glucose-6-phosphate dehydrogenase gene lead stoinactivation of kudna binding during oxidative stress, J Biol Chem., 277,9929-35,2002. [9]. Turanov, A., Hatfıeld, D.L., Gladyshev, V.N., Characterization of Protein Targets of Mammalian Thioredoxin Reductase, Methods enzymol, 474,245-254,2010. [10]. Nishinaka ,Y, Nakamura, H., Masutani, H., Yodoi, J., Redox control of cellular function by thioredoxin: a new therapeuticdirection in host defence, Arch Immunol Their Exp., 49,285-92,2001. [11]. Arner, E.S.J., Nordberg, J. And Holmgren, A., Efncien treduction of lipoamide and lipoicacid by mammalian thioredoxin reductase, Biochem. Biophys. Res. Commun., 225,268-274,1996. [12]. Björnstedt, M., Hamberg, M., Kumar, S., Xue, J.and Holmgren, A., Human thio redoxin reduces lipid hydro peroxides by NADPH and seleno cysteine strongly stimulates there action via catalytically generated selenols, J. Biol. Chem., 270, 11761-11764, 1995. [13]. Arner, E.S.J. and Holmgren, A., Physiological fUnctions of thioredoxin and thioredoxin reductase, Eur. J. Biochem., 267, 6102-6109, 2000. [14]. Holmgren, A., Reduction of disulfides by fhioredoxin. Exceptional reactivity of insülin and suggested functions of thioredoxin in mechanism of hormoneaction, Eur. J. Biochem., 254, 9113-9119,1979. [15]. Rigobello, M.P., Callegaro, M.T., Barzon, E., Benetti, M. and Bindoli, A., Purification of mitochondrial thioredoxin reductase and its involvement in the regulation of membran permeability, Free Radical Biology-Medicine, 24, 370-376, 1998. [16]. May, J.M., Mendiratta, S., Hill, K.E.and Burk, R.F., Reduction of dehydro ascorbate to ascorbate by the selenoenzyme fhioredoxin reductase, J. Biol. Chem., 272, 22607-22610, 1997. [17]. May, J.M., Cobb, C.E., Mendiratta, S., Hill, K.E., Burk, R.F., Reduction of the ascorbyl free radical to ascorbate by thioredoxin reductase, J. Biol. Chem., 273, 23039-23045, 1998. [18]. Jordan, A., Reichard, P. Ribonucleotide reductases. Annual review of biochemistry, 67(1), 71-98, 1998. [19]. Bradford, M.M., Arapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Anal Biochem, 72, 248-251, 1976. [20]. Laemmli, U.K, Cleavage of structural proteins during fhe assembly of the head of bacteriophage T4. Nature, 227, 680-685, 1970. [21]. Baker, A., Payne, C.M., Briehi, M.M. and Powis, G.,Thioredoxin a gene found over expressed in human cancer, mhibits apoptosis invitro and in vivo, Cancer Res., 57(22), 5162-5167, 1997. [22]. Thelander, L., Thioredoxin reductase. Characterization of a homogenous preparation from Escherichia coli B. J Biol. Chem. M, 242(5), 852-859, 1967. [23]. Luthman, M., Holmgren, A., Rat liver thioredoxin and thioredoxin reductase: purification and characterization, Biochemistry, 21, 6628-6633, 1982. [24]. Holmgren, A., Bovine thioredoxin system. Purification of thioredoxin reductase from calf liver and fhymus and studies of its function in disulfide reduction, J Biol. Chem., 252, 4600-4606, 1977. [25]. Gonzales, P.P., Baldesten, A., Reichard, P, Purification of a thioredoxin system from yeast, Journal of Biochem., 245, 2363-2370, 1970. [26]. Tanaka, R. Kosugi, K, Mizukami, M., Ishibashi, M., Tokunaga, H. and Tokunagaa, M. Expression and purification of thioredoxin and thioredoxin reductase from Brevibacillus choshinensis, Protein Expression and Purification, 37, 385-391.2004. [27]. Dai, C., Han, W., Wang, M. H. Two Highly Homologous Methionine Sulfoxide Reductase A from Tomato (Solanum lycopersicum), Exhibit Distinct Catalytic Properties. The Protein Journal, 31(4), 285-292, 2012. [28]. Tandogan, B., Ulusu, N. N. Thioredoxin reductase, Hacettepe J. Biol, Chem, 39, 87-92, 2011. [29]. Carvalho, C, Et. Al, Biomarkers of adverse response to mercury: histopathology versus thioredoxin reductase activity, Journal of Biomedicine and Biotechnology, 2012,1-9, 2012. [30]. Wittle, A.B., Anestal, K., Jerremalm, E., Ehrsson, H., Arner, E.S.J., Inhibition of trxr but not of gr by fhe majör classes of alkylating and platinium-containing anticancer compounds, Free Radical Biology Medicine,39,696-703,2005. [31]. Kemerdere, R. Glial tümörlerde tiyoredoksin redüktaz dengeleri , İstanbul Üniversitesi Cerrah Paşa Tıp Fakültesi Nöroşirürji ABD, İstanbul, 2008.
Yıl 2014,
Cilt: 2 Sayı: 2, 296 - 302, 01.12.2014
Öz
Kaynakça
- [1]. Arner, E. S. and Holmgren, A., Physiological functions of thioredoxin and thioredoxin reductase, Eur. J. Biochem., 267, 6102-6109, 2000. [2]. Söderberg, A., Sahaf, B., Rosen, A., Thioredoxin reductase, a redox active selenoprotein, is secreted by normal and neoplastic cells: presence in human plasma, Cancer research, 60, 2281-2289, 2000. [3]. Lincoln, D.T., Emadi, E.M., Tonissen, K.F., Clarke FM. Thethioredoxin-thioredoxin reductase system: over-expression inhuman cancer, Anticancer Res., 23(3b), 2425-33,2003. [4]. Williams Jr, C. H. Lipoamide dehydrogenase, glutathione reductase, thioredoxin reductase, and mercuric ion reductase a family of flavoenzyme transhydrogenases. Chemistry and biochemistry of flavoenzymes, 3, 121-211, 1992.
- [5]. Powis, G., Montfort, W.R, Properties and biological activities of thioredoxin, Pharmacol Toxicol, 41, 261 -295, 2001. [6]. Mustacich, D., Powis, G., Thioredoxin reductese. BiochemJ. 346,1-8,2000. [7]. Gallegos, A., Berggren, M.I., Gasdaska, J.R. and Powis, G., Mechanisms of the regulation of thioredoxin reductase activity in cancer cells by the chemo preventive agent selenium, Cancer Res., 57,4965-4970,1997. [8]. Ayene, I.S., Stamato, T.D., Mauldin, S.K., Biaglow, J.E., Tuttle, S.W., Jenkins, S.F, Mutation in the glucose-6-phosphate dehydrogenase gene lead stoinactivation of kudna binding during oxidative stress, J Biol Chem., 277,9929-35,2002. [9]. Turanov, A., Hatfıeld, D.L., Gladyshev, V.N., Characterization of Protein Targets of Mammalian Thioredoxin Reductase, Methods enzymol, 474,245-254,2010. [10]. Nishinaka ,Y, Nakamura, H., Masutani, H., Yodoi, J., Redox control of cellular function by thioredoxin: a new therapeuticdirection in host defence, Arch Immunol Their Exp., 49,285-92,2001. [11]. Arner, E.S.J., Nordberg, J. And Holmgren, A., Efncien treduction of lipoamide and lipoicacid by mammalian thioredoxin reductase, Biochem. Biophys. Res. Commun., 225,268-274,1996. [12]. Björnstedt, M., Hamberg, M., Kumar, S., Xue, J.and Holmgren, A., Human thio redoxin reduces lipid hydro peroxides by NADPH and seleno cysteine strongly stimulates there action via catalytically generated selenols, J. Biol. Chem., 270, 11761-11764, 1995. [13]. Arner, E.S.J. and Holmgren, A., Physiological fUnctions of thioredoxin and thioredoxin reductase, Eur. J. Biochem., 267, 6102-6109, 2000. [14]. Holmgren, A., Reduction of disulfides by fhioredoxin. Exceptional reactivity of insülin and suggested functions of thioredoxin in mechanism of hormoneaction, Eur. J. Biochem., 254, 9113-9119,1979. [15]. Rigobello, M.P., Callegaro, M.T., Barzon, E., Benetti, M. and Bindoli, A., Purification of mitochondrial thioredoxin reductase and its involvement in the regulation of membran permeability, Free Radical Biology-Medicine, 24, 370-376, 1998. [16]. May, J.M., Mendiratta, S., Hill, K.E.and Burk, R.F., Reduction of dehydro ascorbate to ascorbate by the selenoenzyme fhioredoxin reductase, J. Biol. Chem., 272, 22607-22610, 1997. [17]. May, J.M., Cobb, C.E., Mendiratta, S., Hill, K.E., Burk, R.F., Reduction of the ascorbyl free radical to ascorbate by thioredoxin reductase, J. Biol. Chem., 273, 23039-23045, 1998. [18]. Jordan, A., Reichard, P. Ribonucleotide reductases. Annual review of biochemistry, 67(1), 71-98, 1998. [19]. Bradford, M.M., Arapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Anal Biochem, 72, 248-251, 1976. [20]. Laemmli, U.K, Cleavage of structural proteins during fhe assembly of the head of bacteriophage T4. Nature, 227, 680-685, 1970. [21]. Baker, A., Payne, C.M., Briehi, M.M. and Powis, G.,Thioredoxin a gene found over expressed in human cancer, mhibits apoptosis invitro and in vivo, Cancer Res., 57(22), 5162-5167, 1997. [22]. Thelander, L., Thioredoxin reductase. Characterization of a homogenous preparation from Escherichia coli B. J Biol. Chem. M, 242(5), 852-859, 1967. [23]. Luthman, M., Holmgren, A., Rat liver thioredoxin and thioredoxin reductase: purification and characterization, Biochemistry, 21, 6628-6633, 1982. [24]. Holmgren, A., Bovine thioredoxin system. Purification of thioredoxin reductase from calf liver and fhymus and studies of its function in disulfide reduction, J Biol. Chem., 252, 4600-4606, 1977. [25]. Gonzales, P.P., Baldesten, A., Reichard, P, Purification of a thioredoxin system from yeast, Journal of Biochem., 245, 2363-2370, 1970. [26]. Tanaka, R. Kosugi, K, Mizukami, M., Ishibashi, M., Tokunaga, H. and Tokunagaa, M. Expression and purification of thioredoxin and thioredoxin reductase from Brevibacillus choshinensis, Protein Expression and Purification, 37, 385-391.2004. [27]. Dai, C., Han, W., Wang, M. H. Two Highly Homologous Methionine Sulfoxide Reductase A from Tomato (Solanum lycopersicum), Exhibit Distinct Catalytic Properties. The Protein Journal, 31(4), 285-292, 2012. [28]. Tandogan, B., Ulusu, N. N. Thioredoxin reductase, Hacettepe J. Biol, Chem, 39, 87-92, 2011. [29]. Carvalho, C, Et. Al, Biomarkers of adverse response to mercury: histopathology versus thioredoxin reductase activity, Journal of Biomedicine and Biotechnology, 2012,1-9, 2012. [30]. Wittle, A.B., Anestal, K., Jerremalm, E., Ehrsson, H., Arner, E.S.J., Inhibition of trxr but not of gr by fhe majör classes of alkylating and platinium-containing anticancer compounds, Free Radical Biology Medicine,39,696-703,2005. [31]. Kemerdere, R. Glial tümörlerde tiyoredoksin redüktaz dengeleri , İstanbul Üniversitesi Cerrah Paşa Tıp Fakültesi Nöroşirürji ABD, İstanbul, 2008.
Toplam 2 adet kaynakça vardır.
Ayrıntılar
| Bölüm | Araştırma Makalesi |
|---|---|
| Yazarlar | |
| Yayımlanma Tarihi | 1 Aralık 2014 |
| Yayımlandığı Sayı | Yıl 2014 Cilt: 2 Sayı: 2 |
