Afinite Kromatografisi Yöntemi ile Saflaştırılan Karbonik Anhidraz (I-II) İzoenzimleri Üzerine Bazı Pestisitlerin İn Vitro Etkilerinin Araştırılması

Öz

Bu çalışmanın amacı; çevre ve insan sağlığı için toksisitesi yüksek olan bazı pestisitlerin hidrataz aktivite yöntemi kullanılarak karbonik anhidraz izoenzimleri (hCA I ve II) üzerine etkilerini araştırmaktır. İnsan kan eritrositlerinden Sefaroz-4B-4-(6-Amino-heksilokzi)-benzensülfonamit afinite jeli kullanılarak hCA I ve II izoenzimleri saflaştırıldı. Enzimlerin saflığı; sodyum dodesil sülfat poliakrilamid jel elektroforezi ile kontrol edildi. Ayrıca bazı pestisitlerin hCAI-II izoenzimlerini in vitro ortamda µM düzeylerde inhibe ettiği belirlendi. Bu pestisitlerden hCA-I enzimi için en güçlü inhibitörün Simazin (IC50: 0,366 µM), en zayıf ise Koumatetralil (IC50: 6,41 µM) olduğu belirlendi. hCA-II enzimi için en güçlü inhibitörün Klorpirifos (IC50: 0,527 µM), en zayıf inhibitörün ise Koumatetralil (IC50: 8,74 µM) olduğu belirlendi.

Anahtar Kelimeler

• Karbonik anhidraz
• Pestisitler
• Enzim inhibisyonu

Investigation of In Vitro Effects of Some Pesticides on Carbonic Anhydrase (I-II) Isoenzymes Purified by Affinity Chromatography Method

Öz

The aim of this study was to investigate the effects of some pesticides with high toxicity for the environment and human health on carbonic anhydrase isoenzymes (hCA I and II) using hydratase activity method. hCA I-II were purified from human blood erythrocytes using Sepharose-4B-4-(6-Amino-hexyloxy)-benzenesulfonamide affinity gel. In order to determine the purity of the enzymes, sodium dedocyl sulphate polyacrylamide gel electrophoresis was performed. In addition, it was determined that some pesticides inhibited hCAI-II isoenzymes at µM levels in vitro. It was determined that among these pesticides, the strongest inhibitor for the hCA-I enzyme was simazine (IC50: 0.366 µM), and the weakest was coumatetralil (IC50: 6.41 µM). The strongest inhibitor for the hCA- II enzyme was chlorpyrifos (IC50: 0.527 µM), and the weakest inhibitor was coumatetralil (IC50: 8.74 µM).

Anahtar Kelimeler

• Carbonic anhydrase
• Pesticides Enzyme inhibition

Kaynakça

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