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Purification and partial characterization of thioredoxin reductase from the hepatopancreas of the mollusc Mytilus galloprovincialis Lam.

Year 2018, Volume 48, Issue 1, 12 - 17, 30.04.2018

Abstract

DOI: 10.5152/IstanbulJPharm.2018.382627


Thioredoxin reductase (TrxR, EC 1.6.4.5) is a ubiquitous flavoenzyme that is present from Archaea to humans, and it is the only enzyme capable of catalyzing the reduction of thioredoxin (Trx) by nicotinamide adenine dinucleotide phosphate (NADPH). Although TrxR has been purified and characterized from different bacteria, plants, and mammalian organisms, a survey of the literature revealed no studies on the purification and characterization of TrxR from the mussel Mytilus galloprovincialis Lam. In this study, TrxR was purified to homogeneity from the hepatopancreatic tissue of M. galloprovincialis Lam. by extraction, ammonium sulfate precipitation, and DEAE-Sepharose CL-6B anion and 2’,5‘-ADP-agarose chromatographies, and some of its kinetic properties were examined. Molar mass determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed only a single protein band corresponding to a molecular weight of 35 kDa. Optimum pH and temperature were found to be 7.0 and 60°C, respectively. Km and Vmax values for NADPH were found to be 85 µmol and 4.82 µmol/min/mg, respectively. For 5,5ʹ-dithiobis (2-nitrobenzoic) acid (DTNB), the Km and Vmax values were 193 µmol and 1.32 µmol/min/mg, respectively. Increasing the knowledge on the kinetic properties of TrxR will significantly increase the prospects of enzyme application as an oxidative stress biomarker in mussels and fishes for monitoring contamination in coastal environments.

References

  • Arısan-Ataç İ, Kuşçu İ, Berkkan H (1994). α-D-Glucosidase activities in the mantle tissue of the mussel Mytilus galloprovincialis L. Acta Pharmaceutica Turcica 36: 53-58.
  • Arnér ES, Holmgren A (2000). Measurement of thioredoxin and thioredoxin reductase. Curr Protoc Toxicol 24(7.4): 7.4.1-7.4.14.
  • Arnér ESJ, Holmgren A (2006). The thioredoxin system in cancer. Semin Cancer Biol 16(6): 420-426.
  • Arias DG, Regner EL, Iglesias AA, Guerrero SA (2010). Entamoeba histolytica thioredoxin reductase: molecular and functional characterization of its atypical properties. Biochim Biophys Acta 1820(12): 1859-1866.
  • Bradford MM (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72(1-2): 248-254.
  • Brodelius P, Larsson PO, Mosbach K (1974). The Synthesis of three AMP-analogues: N6-(6-aminohexy1)-adenosine 5’-monophosphate, N6-(6-aminohexyl)-adenosine 2’,5’-bisphosphate, and N6-(6-aminohexyl)-adenosine 3’, 5’-bisphosphate and their application as general ligands in biospecific affinity chromatography. Eur J Biochem 47(1): 81-89.
  • Brown DM, Upcroft JA, Upcroft P (1996). A thioredoxin reductase-class of disulphide reductase in the protozoan parasite Giardia duodenalis. Mol Biochem Parasitol 83(2): 211-220.
  • Can A, Akev N, Askhasi A (2000). Partial purification and characterization of aspartate aminotransferase from the hepatopancreas of the mussel Mytilus galloprovincialis Lam. J Fac Pharm Istanbul 33: 37-48.
  • Coombs GH, Westrop GD, Suchan P, Puzova G, Hirt RP, Embley TM, Mottram JC, Müller S (2004). The amitochondriate eukaryote Trichomonas vaginalis contains a divergent thioredoxin-linked peroxiredoxin antioxidant system. J Biol Chem 279(7): 5249-5256.
  • Dönmez Yurtpınar Ö, Yılmaz Özden T, Can A (2014). Partial purification and some properties of aspartate aminotransferase from the mantle tissue of Mytilus galloprovincialis Lam. J Fac Pharm Istanbul 44(1): 1-10.
  • Gromer S, Arscott LD, Williams CHJr, Schirmer RH (1998). Human placenta thioredoxin reductase. J Biol Chem 273: 20096-20101.
  • Harms C, Meyer MA, Andreesen JR (1998). Fast purification of thioredoxin reductases and of thioredoxins with an unusual redox-active centre from anaerobic, amino-acid-utilizing bacteria. Microbiology 144(3): 793-800.
  • Holmgren A, Bjorsnstedt M (1995). Thioredoxin and thioredoxin reductase. Methods Enzymol 252: 199-208.
  • Horecká T, Perecko D, Kutejová E, Mikuláŝová D, Kollárová M (1998). Purification and partial characterization of thioredoxin reductase from Streptomyces aureofaciens. Biochem Mol Biol Int 46(4): 657-665.
  • Kapoor G, Banyal HS (2011). Purification and characterization of Plasmodium berghei thioredoxin reductase. Asian J Anim Sci 5(2): 145-152.
  • Laemmli UK (1970). Cleavage of structural proteins during the assembly of the head bacteriophage T4. Nature 227(5259): 680-685.
  • Larsson A (1973). Thioredoxin reductase from rat liver. Eur J Biochem 35(2): 346-349.
  • Lu J, Zhoong L, Lönn ME, Burk RF, Hill KE, Holmgren A (2009). Penultimate selenocysteine residue replaced by cysteine in thioredoxin reductase from selenium-deficient rat liver. FASEB J 23(8): 2394-2402.
  • Luthman M, Holmgren A (1982). Rat liver thioredoxin and thioredoxin reductase: purification and characterization. Biochemistry 21(26): 6628-6633.
  • Maggioli G, Piacenza L, Carambula B, Carmona C (2004). Purification, characterization, and immunolocalization of a thioredoxin reductase from adult Fasciola hepatica. J Parasitol 90(2): 205-211.
  • Moore EC, Reichard P, Thelander L (1964). Enzymatic synthesis of deoxyribonucleotides: V. Purification and properties of thioredoxin reductase from Escherichia coli B. J Biol Chem 239(10): 3445-3452.
  • Özgençli İ, Çiftçi M (2016). Purification and characterization of mitochondrial thioredoxin reductase enzyme from rainbow trout (Oncorhynchus mykiss) liver and investigation of the in vitro effects of some metal ions on the enzyme. Turk J Chem 40(1): 174-183.
  • Özsoy N, Berkkan H (1997). β-Glucosidase activities in the mantle tissue of the mussel Mytilus galloprovincialis L. Acta Pharmaceutica Turcica 39(3): 99-103.
  • Pigiet VP, Conley RR (1977). Purification of thioredoxin, thioredoxine reductase and glutathione reductase by affinity chromatography. J Biol Chem 252(18): 6367-6372.
  • Reichheld JP, Meyer E, Khafif M, Bonnard G, Meyer Y (2005). AtNTRB is the majör mitochondrial thioredoxin reductase in Arabidopsis thaliana. FEBS Lett 579(2): 337-342.
  • Seo HJ, Lee YN (2010). Characterization of Deinococcus radiophilus thioredoxin reductase active with both NADH and NADPH. J Microbiol 48(5): 637-643.
  • Somar N, Özsoy N, Berkkan H (2000). b-Galactosidase activity in the mantle tissue of the mussel Mytilus galloprovincialis Lam. Acta Pharmaceutica Turcica 42(2,3): 80-84.
  • Sureda A, Box A, Tejada S, Blanco A, Caixach J, Deudero S (2011). Biochemical responses of Mytilus galloprovincialis as biomarkers of acute environmental pollution caused by Don Pedro oil spill (Eivissa Island, Spain). Aquat Toxicol 101(3): 540-549.
  • Yalvaç S, Kuşçu İ (1993). β-D-Galactosidase isoenzymes of Mytilus galloprovincialis L. Acta Pharmaceutica Turcica 35: 45-50.
  • Watabe S, Makino Y, Ogawa K, Hiroi T, Yamamoto Y, Takahashi SY (1999). Mitochondrial thioredoxin reductase in bovine adrenal cortex. Eur J Biochem 264(1): 74-84.
  • Williams CH, Zanetti G, Arscott LD, McAllister JK (1967). Lypoamide dehydrogenase, glutathione reductase, thioredoxin reductase, and thioredoxin. J Biol Chem 242(22): 5226-5231.
  • Williams CH (1995). Mechanism and structure of thioredoxin reductase from Escherichia coli. FASEB J 9(13): 1267-1276.
  • Williams CH, Arscott LD, Müller S, Lennon BW, Ludwig ML, Wang P-F, Veine DM, Becker K, Schirmer RH (2000). Thioredoxin reductase. Two modes of catalysis have evolved. Minireview. Eur J Biochem 267: 6110-6117.
  • Yang X, Ma K (2010). Characterization of a thioredoxin-thioredoxin reductase system from the hyperthermophilic bacterium Thermotoga maritima. J Bacteriol 192(5): 1370-1376.

Year 2018, Volume 48, Issue 1, 12 - 17, 30.04.2018

Abstract

References

  • Arısan-Ataç İ, Kuşçu İ, Berkkan H (1994). α-D-Glucosidase activities in the mantle tissue of the mussel Mytilus galloprovincialis L. Acta Pharmaceutica Turcica 36: 53-58.
  • Arnér ES, Holmgren A (2000). Measurement of thioredoxin and thioredoxin reductase. Curr Protoc Toxicol 24(7.4): 7.4.1-7.4.14.
  • Arnér ESJ, Holmgren A (2006). The thioredoxin system in cancer. Semin Cancer Biol 16(6): 420-426.
  • Arias DG, Regner EL, Iglesias AA, Guerrero SA (2010). Entamoeba histolytica thioredoxin reductase: molecular and functional characterization of its atypical properties. Biochim Biophys Acta 1820(12): 1859-1866.
  • Bradford MM (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72(1-2): 248-254.
  • Brodelius P, Larsson PO, Mosbach K (1974). The Synthesis of three AMP-analogues: N6-(6-aminohexy1)-adenosine 5’-monophosphate, N6-(6-aminohexyl)-adenosine 2’,5’-bisphosphate, and N6-(6-aminohexyl)-adenosine 3’, 5’-bisphosphate and their application as general ligands in biospecific affinity chromatography. Eur J Biochem 47(1): 81-89.
  • Brown DM, Upcroft JA, Upcroft P (1996). A thioredoxin reductase-class of disulphide reductase in the protozoan parasite Giardia duodenalis. Mol Biochem Parasitol 83(2): 211-220.
  • Can A, Akev N, Askhasi A (2000). Partial purification and characterization of aspartate aminotransferase from the hepatopancreas of the mussel Mytilus galloprovincialis Lam. J Fac Pharm Istanbul 33: 37-48.
  • Coombs GH, Westrop GD, Suchan P, Puzova G, Hirt RP, Embley TM, Mottram JC, Müller S (2004). The amitochondriate eukaryote Trichomonas vaginalis contains a divergent thioredoxin-linked peroxiredoxin antioxidant system. J Biol Chem 279(7): 5249-5256.
  • Dönmez Yurtpınar Ö, Yılmaz Özden T, Can A (2014). Partial purification and some properties of aspartate aminotransferase from the mantle tissue of Mytilus galloprovincialis Lam. J Fac Pharm Istanbul 44(1): 1-10.
  • Gromer S, Arscott LD, Williams CHJr, Schirmer RH (1998). Human placenta thioredoxin reductase. J Biol Chem 273: 20096-20101.
  • Harms C, Meyer MA, Andreesen JR (1998). Fast purification of thioredoxin reductases and of thioredoxins with an unusual redox-active centre from anaerobic, amino-acid-utilizing bacteria. Microbiology 144(3): 793-800.
  • Holmgren A, Bjorsnstedt M (1995). Thioredoxin and thioredoxin reductase. Methods Enzymol 252: 199-208.
  • Horecká T, Perecko D, Kutejová E, Mikuláŝová D, Kollárová M (1998). Purification and partial characterization of thioredoxin reductase from Streptomyces aureofaciens. Biochem Mol Biol Int 46(4): 657-665.
  • Kapoor G, Banyal HS (2011). Purification and characterization of Plasmodium berghei thioredoxin reductase. Asian J Anim Sci 5(2): 145-152.
  • Laemmli UK (1970). Cleavage of structural proteins during the assembly of the head bacteriophage T4. Nature 227(5259): 680-685.
  • Larsson A (1973). Thioredoxin reductase from rat liver. Eur J Biochem 35(2): 346-349.
  • Lu J, Zhoong L, Lönn ME, Burk RF, Hill KE, Holmgren A (2009). Penultimate selenocysteine residue replaced by cysteine in thioredoxin reductase from selenium-deficient rat liver. FASEB J 23(8): 2394-2402.
  • Luthman M, Holmgren A (1982). Rat liver thioredoxin and thioredoxin reductase: purification and characterization. Biochemistry 21(26): 6628-6633.
  • Maggioli G, Piacenza L, Carambula B, Carmona C (2004). Purification, characterization, and immunolocalization of a thioredoxin reductase from adult Fasciola hepatica. J Parasitol 90(2): 205-211.
  • Moore EC, Reichard P, Thelander L (1964). Enzymatic synthesis of deoxyribonucleotides: V. Purification and properties of thioredoxin reductase from Escherichia coli B. J Biol Chem 239(10): 3445-3452.
  • Özgençli İ, Çiftçi M (2016). Purification and characterization of mitochondrial thioredoxin reductase enzyme from rainbow trout (Oncorhynchus mykiss) liver and investigation of the in vitro effects of some metal ions on the enzyme. Turk J Chem 40(1): 174-183.
  • Özsoy N, Berkkan H (1997). β-Glucosidase activities in the mantle tissue of the mussel Mytilus galloprovincialis L. Acta Pharmaceutica Turcica 39(3): 99-103.
  • Pigiet VP, Conley RR (1977). Purification of thioredoxin, thioredoxine reductase and glutathione reductase by affinity chromatography. J Biol Chem 252(18): 6367-6372.
  • Reichheld JP, Meyer E, Khafif M, Bonnard G, Meyer Y (2005). AtNTRB is the majör mitochondrial thioredoxin reductase in Arabidopsis thaliana. FEBS Lett 579(2): 337-342.
  • Seo HJ, Lee YN (2010). Characterization of Deinococcus radiophilus thioredoxin reductase active with both NADH and NADPH. J Microbiol 48(5): 637-643.
  • Somar N, Özsoy N, Berkkan H (2000). b-Galactosidase activity in the mantle tissue of the mussel Mytilus galloprovincialis Lam. Acta Pharmaceutica Turcica 42(2,3): 80-84.
  • Sureda A, Box A, Tejada S, Blanco A, Caixach J, Deudero S (2011). Biochemical responses of Mytilus galloprovincialis as biomarkers of acute environmental pollution caused by Don Pedro oil spill (Eivissa Island, Spain). Aquat Toxicol 101(3): 540-549.
  • Yalvaç S, Kuşçu İ (1993). β-D-Galactosidase isoenzymes of Mytilus galloprovincialis L. Acta Pharmaceutica Turcica 35: 45-50.
  • Watabe S, Makino Y, Ogawa K, Hiroi T, Yamamoto Y, Takahashi SY (1999). Mitochondrial thioredoxin reductase in bovine adrenal cortex. Eur J Biochem 264(1): 74-84.
  • Williams CH, Zanetti G, Arscott LD, McAllister JK (1967). Lypoamide dehydrogenase, glutathione reductase, thioredoxin reductase, and thioredoxin. J Biol Chem 242(22): 5226-5231.
  • Williams CH (1995). Mechanism and structure of thioredoxin reductase from Escherichia coli. FASEB J 9(13): 1267-1276.
  • Williams CH, Arscott LD, Müller S, Lennon BW, Ludwig ML, Wang P-F, Veine DM, Becker K, Schirmer RH (2000). Thioredoxin reductase. Two modes of catalysis have evolved. Minireview. Eur J Biochem 267: 6110-6117.
  • Yang X, Ma K (2010). Characterization of a thioredoxin-thioredoxin reductase system from the hyperthermophilic bacterium Thermotoga maritima. J Bacteriol 192(5): 1370-1376.

Details

Primary Language English
Subjects Pharmacology and Pharmacy
Journal Section Original Article
Authors

Esra ACAR This is me
Department of Biochemistry, Faculty of Medicine, Kocaeli University, 41380, Kocaeli, Turkey
0000-0002-0814-0820


Gözde HASBAL This is me
Department of Biochemistry, Faculty of Pharmacy, Istanbul University, 34116, Istanbul, Turkey
0000-0002-0216-7635


Nurten ÖZSOY> (Primary Author)
Department of Biochemistry, Faculty of Pharmacy, Istanbul University, 34116, Istanbul, Turkey
0000-0002-2419-9128

Publication Date April 30, 2018
Application Date January 23, 2018
Acceptance Date
Published in Issue Year 2018, Volume 48, Issue 1

Cite

Bibtex @research article { iujp419900, journal = {İstanbul Journal of Pharmacy}, eissn = {2587-2087}, address = {İstanbul Üniversitesi Eczacılık Fakültesi Farmasötik Botanik AD}, publisher = {Istanbul University}, year = {2018}, volume = {48}, number = {1}, pages = {12 - 17}, title = {Purification and partial characterization of thioredoxin reductase from the hepatopancreas of the mollusc Mytilus galloprovincialis Lam.}, key = {cite}, author = {Acar, Esra and Hasbal, Gözde and Özsoy, Nurten} }
APA Acar, E. , Hasbal, G. & Özsoy, N. (2018). Purification and partial characterization of thioredoxin reductase from the hepatopancreas of the mollusc Mytilus galloprovincialis Lam. . İstanbul Journal of Pharmacy , 48 (1) , 12-17 . Retrieved from https://dergipark.org.tr/en/pub/iujp/issue/36850/419900
MLA Acar, E. , Hasbal, G. , Özsoy, N. "Purification and partial characterization of thioredoxin reductase from the hepatopancreas of the mollusc Mytilus galloprovincialis Lam." . İstanbul Journal of Pharmacy 48 (2018 ): 12-17 <https://dergipark.org.tr/en/pub/iujp/issue/36850/419900>
Chicago Acar, E. , Hasbal, G. , Özsoy, N. "Purification and partial characterization of thioredoxin reductase from the hepatopancreas of the mollusc Mytilus galloprovincialis Lam.". İstanbul Journal of Pharmacy 48 (2018 ): 12-17
RIS TY - JOUR T1 - Purification and partial characterization of thioredoxin reductase from the hepatopancreas of the mollusc Mytilus galloprovincialis Lam. AU - EsraAcar, GözdeHasbal, NurtenÖzsoy Y1 - 2018 PY - 2018 N1 - DO - T2 - İstanbul Journal of Pharmacy JF - Journal JO - JOR SP - 12 EP - 17 VL - 48 IS - 1 SN - -2587-2087 M3 - UR - Y2 - 2022 ER -
EndNote %0 İstanbul Journal of Pharmacy Purification and partial characterization of thioredoxin reductase from the hepatopancreas of the mollusc Mytilus galloprovincialis Lam. %A Esra Acar , Gözde Hasbal , Nurten Özsoy %T Purification and partial characterization of thioredoxin reductase from the hepatopancreas of the mollusc Mytilus galloprovincialis Lam. %D 2018 %J İstanbul Journal of Pharmacy %P -2587-2087 %V 48 %N 1 %R %U
ISNAD Acar, Esra , Hasbal, Gözde , Özsoy, Nurten . "Purification and partial characterization of thioredoxin reductase from the hepatopancreas of the mollusc Mytilus galloprovincialis Lam.". İstanbul Journal of Pharmacy 48 / 1 (April 2018): 12-17 .
AMA Acar E. , Hasbal G. , Özsoy N. Purification and partial characterization of thioredoxin reductase from the hepatopancreas of the mollusc Mytilus galloprovincialis Lam.. iujp. 2018; 48(1): 12-17.
Vancouver Acar E. , Hasbal G. , Özsoy N. Purification and partial characterization of thioredoxin reductase from the hepatopancreas of the mollusc Mytilus galloprovincialis Lam.. İstanbul Journal of Pharmacy. 2018; 48(1): 12-17.
IEEE E. Acar , G. Hasbal and N. Özsoy , "Purification and partial characterization of thioredoxin reductase from the hepatopancreas of the mollusc Mytilus galloprovincialis Lam.", İstanbul Journal of Pharmacy, vol. 48, no. 1, pp. 12-17, Apr. 2018