PON1 enzimi üzerine DT-15-Crown-5 taç eterinin etkisinin araştırılması

İnsan serum paraoksonaz 1 (PON1, EC3.1.8.1) HDL'ye bağlı olan, aktif bölgesinde kalsiyum içeren, organofosfat ajanlarını ve sinir gazlarını hidroliz eden, LDL'nin oksidasyonu ile lipit peroksitlerin oluşumuna ve bakteri endotoksinlerine karşı koruyucu etkisi olan önemli bir karaciğer enzimidir. Çalışmamızda, PON1 enzimi daha önceden sentezi yapılmış, sepharose-4B-L-tirosin-1-naftilamin jeli kullanılarak hidrofobik etkileşim kromatografisi ile tamamen saflaştırılmıştır. Enzimin saflık kontrolü de SDS – PAGE ile yapılmıştır. İnhibisyon ajanı olarak kullanılmak üzere 7,13-dithio-1,4,10-trioksasiklopentadekan (DT-15-Crown-5) tiyo taç eteri SN2 mekanizmasına göre halka kapama reaksiyonu sonucunda sentezlendi. DT-15-Crown-5’in paraoksanaz enzimi üzerinde kuvvetli inhibisyon etkisi gözlendi. DT-Taçeterin PON1 ile inhibisyon değeri (IC50) 22,62 M olarak tespit edildi.

Investigation of the effect of DT-15-Crown-5 crown ether on PON1 enzyme

Human serum paraoxonase 1 (PON1, EC3.1.8.1) is an important liver enzyme which is bound to HDL, contains calcium in its active site, hydrolyses organophosphate agents and nerve gases, has protective effect against the formation of lipid peroxides by oxidation of LDL and bacterial endotoxins. In our study, in order to purify the enzyme PON1, it was purified by hydrophobic interaction chromatography using sepharose-4B-L-tyrosine-1-naphthylamine gel. The purity of the enzyme was also checked by SDS - PAGE. 7,13-Dithio-1,4,10-trioxacyclopentadecane (DT-15-Crown-5) thio crown ether was synthesized as a result of the ring closure reaction according to the SN2 mechanism to be used as an inhibiting agent. Strong inhibition effect of DT-15-Crown-5 on paraoxanase enzyme was observed. Inhibition value (IC50) of DT-Taçeter with PON1 was determined as 22.62 M.

Keywords:

Thiocrown ether, paraoxanase (PON) inhibition, hydrophobic interaction chromatography,

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