SavedAdd your journal

Journals / İstanbul Journal of Pharmacy / 2018 / Cilt: 48 - Sayı: 1

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

Esra ACAR
Gözde HASBAL
Nurten ÖZSOY
Pages
12–17
Publication Date
2018-04-30
DOI
—
🏛️Bibliographic metadata and full text access for this article are provided via DergiPark.
DergiPark Record ↗
✨

AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

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.

📌 3 Key Takeaways

🎯
Objective & Hypothesis:DOI: 10.5152/IstanbulJPharm.2018.382627Thioredoxin 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).
📊
Key Findings & Data: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.
💡
Conclusion & Impact: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.
🔬Design:Survey & Empirical Study
📍Scope:2018 Verileri
⚙️Variables:Enzyme purification · kinetic properties · Mytilus galloprovincialis Lam.

Abstract

DOI: 10.5152/IstanbulJPharm.2018.382627Thioredoxin 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.

🔖

Cite This Article

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, Cilt: 48 - Sayı: 1, 12–17. https://ajindex.com/articles/41777

📄 Full Text PDF Viewer

📄

PDF görüntüleyici hazırlanıyor...

References (11)

  1. 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.
  2. 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.
  3. 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.
  4. 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.
  5. 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.
  6. 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.
  7. 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.
  8. 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.
  9. Gromer S, Arscott LD, Williams CHJr, Schirmer RH (1998). Human placenta thioredoxin reductase. J Biol Chem 273: 20096-20101.
  10. 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.
  11. 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