Yıl 2017, Cilt 17, Sayı 4, Sayfalar 385 - 394 2017-12-01

In vitro Inhibition Effects of Some Metal Ions on Glutathione Reductase Purified from Capoeta trutta Kidney
Capoeta trutta Böbrek Dokusundan Saflaştırılan Glutatyon Redüktaz Enzim Aktivitesine Bazı Metallerin in vitro İnhibisyon Etkileri

Muammer KIRICI [1] , Mahinur KIRICI [2] , Muhammed ATAMANALP [3]

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Glutathione reductase (EC: 1.8.1.7; GR) was purified from the kidney of the teleost fish Capoeta trutta. The purification procedure consisted of three steps: preparation of homogenate, ammonium sulfate fractionation and affinity chromatography on 2’,5’-ADP Sepharose 4B. The enzyme was purified 794-fold with a yield of 35.4% and a specific activity of 11.91 U/mg proteins. In order to control enzyme purity, SDS-PAGE was done and showed a single band for enzyme. A single band was obtained approximately at 55 kDa. In addition, inhibitory effects of metal ions (Ag+, Co+2, Ni+2, Cu+2 and Zn+2) on fish kidney glutathione reductase were investigated. Ki constants and IC50 values for metal ions were determined by Lineweaver–Burk graphs and plotting activity % vs. [I], respectively. IC50 values were 0.00078, 0.622, 0.722, 0.073 and 0.519 mM, and Ki constants were 0.000394 ± 0.0002, 0.235 ± 0.027, 0.279 ± 0.048, 0.026 ± 0.008 and 0.382 ± 0.024 mM for Ag+, Co+2, Ni+2, Cu+2 and Zn+2, respectively. From these results, we showed that Ag+ is the most potent inhibitor of glutathione reductase enzyme.

Bu çalışmada, glutatyon redüktaz (EC: 1.8.1.7; GR) iç su balıklarından Capoeta trutta böbrek dokusundan saflaştırılmıştır. Saflaştırma işlemi, homojenatın hazırlanması, amonyum sülfat çöktürmesi ve 2’,5’-ADP Sepharose 4B afinite kromatografisi olmak üzere 3 basamakta gerçekleştirilmiştir. Bu yöntemlerle, enzim 11.91 EÜ/mg protein spesifik aktivite ve %35.4 verimle 794 kat saflaştırılmıştır. Saflaştırma sonucu elde edilen saf enzimin saflık kontrolü sodyum dodesilsülfat poliakrilamid jel elektroforezi (SDS-PAGE) ile yapıldı ve enzimlerin tek band olduğu görülmüştür. Enzimin molekül kütlesi yaklaşık olarak 55 kDa olarak hesaplanmıştır. Ayrıca, balık böbrek dokusundan saflaştırılan glutatyon redüktaz enzimi üzerine bazı metallerin (Ag+, Co+2, Ni+2, Cu+2 ve Zn+2) in vitro inhibisyon etkileri incelenmiştir. Metallerin Ki sabiti ve IC50 değerleri sırasıyla Lineweaver-Burk ve Yüzde (%) Aktivite-[Metal] grafikleri ile hesaplanmıştır. Bu grafiklerden, Ag+, Co+2, Ni+2, Cu+2 ve Zn+2 metalleri için IC50 değerleri sırasıyla 0.00078, 0.622, 0.722, 0.073 ve 0.519 mM, ve Ki sabitleri sırasıyla 0.000394 ± 0.0002, 0.235 ± 0.027, 0.279 ± 0.048, 0.026 ± 0.008 ve 0.382 ± 0.024 mM olarak hesaplanmıştır. Sonuç olarak, Ag+ metalinin diğer metallerden daha fazla C. trutta böbrek GR enzimini inhibe ettiği tespit edilmiştir.


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Konular Yaşam Bilimleri
Dergi Bölümü Araştırma Makalesi
Yazarlar

Orcid: orcid.org/0000-0003-1888-4388
Yazar: Muammer KIRICI
E-posta: muammerkirici@hotmail.com
Kurum: Bingöl University, Faculty of Agriculture, Department of Fisheries, Bingöl, Turkey.
Ülke: Turkey


Yazar: Mahinur KIRICI
E-posta: mahinur_cam@hotmail.com
Kurum: Bingöl University, Faculty of Arts and Science, Department of Chemistry, Bingöl, Turkey.
Ülke: Turkey


Yazar: Muhammed ATAMANALP
E-posta: mataman@atauni.edu.tr
Kurum: Atatürk University, Faculty of Fisheries, Department of Aquaculture, Erzurum, Turkey.
Ülke: Turkey


Bibtex @araştırma makalesi { yunusae337640, journal = {Aquaculture Studies (Eski Yunus Araştırma Bülteni)}, issn = {2618-6381}, address = {Su Ürünleri Merkez Araştırma Enstitüsü}, year = {2017}, volume = {17}, pages = {385 - 394}, doi = {10.17693/yunusae.v17i31121.337640}, title = {In vitro Inhibition Effects of Some Metal Ions on Glutathione Reductase Purified from Capoeta trutta Kidney}, key = {cite}, author = {ATAMANALP, Muhammed and KIRICI, Muammer and KIRICI, Mahinur} }
APA KIRICI, M , KIRICI, M , ATAMANALP, M . (2017). In vitro Inhibition Effects of Some Metal Ions on Glutathione Reductase Purified from Capoeta trutta Kidney. Aquaculture Studies (Eski Yunus Araştırma Bülteni), 17 (4), 385-394. DOI: 10.17693/yunusae.v17i31121.337640
MLA KIRICI, M , KIRICI, M , ATAMANALP, M . "In vitro Inhibition Effects of Some Metal Ions on Glutathione Reductase Purified from Capoeta trutta Kidney". Aquaculture Studies (Eski Yunus Araştırma Bülteni) 17 (2017): 385-394 <http://dergipark.gov.tr/yunusae/issue/31121/337640>
Chicago KIRICI, M , KIRICI, M , ATAMANALP, M . "In vitro Inhibition Effects of Some Metal Ions on Glutathione Reductase Purified from Capoeta trutta Kidney". Aquaculture Studies (Eski Yunus Araştırma Bülteni) 17 (2017): 385-394
RIS TY - JOUR T1 - In vitro Inhibition Effects of Some Metal Ions on Glutathione Reductase Purified from Capoeta trutta Kidney AU - Muammer KIRICI , Mahinur KIRICI , Muhammed ATAMANALP Y1 - 2017 PY - 2017 N1 - doi: 10.17693/yunusae.v17i31121.337640 DO - 10.17693/yunusae.v17i31121.337640 T2 - Aquaculture Studies (Eski Yunus Araştırma Bülteni) JF - Journal JO - JOR SP - 385 EP - 394 VL - 17 IS - 4 SN - 2618-6381- M3 - doi: 10.17693/yunusae.v17i31121.337640 UR - http://dx.doi.org/10.17693/yunusae.v17i31121.337640 Y2 - 2017 ER -
EndNote %0 Aquaculture Studies (Eski Yunus Araştırma Bülteni) In vitro Inhibition Effects of Some Metal Ions on Glutathione Reductase Purified from Capoeta trutta Kidney %A Muammer KIRICI , Mahinur KIRICI , Muhammed ATAMANALP %T In vitro Inhibition Effects of Some Metal Ions on Glutathione Reductase Purified from Capoeta trutta Kidney %D 2017 %J Aquaculture Studies (Eski Yunus Araştırma Bülteni) %P 2618-6381- %V 17 %N 4 %R doi: 10.17693/yunusae.v17i31121.337640 %U 10.17693/yunusae.v17i31121.337640