Bazı antrakinonların insan paraoksonaz 1 (hPON1) üzerine etkilerinin incelenmesi

Abstract

Antikanser antibiyotikler olarak yaygın kullanılan bazı dihidroksiantrakinon türevlerinin (antrorufin, ksantopurpirin, krisazin ve krizofanol) saflaştırılmış serum paraoksonaz 1 (PON1) aktivitesi üzerine (EC 3.1.8.1) in vitro etkileri araştırıldı.  Paraoksonaz 1, insan kan serumundan 355,12 kat, % 55 verimle, amonyum sülfat çöktürmesi ve Sepharose 4B-L-tirozin-1-naftilamin hidrofobik etkileşim kromatografisi kullanılarak saflaştırılmıştır. Saflaştırılmış enzimin Sodyum Dodesil Sülfat-poliakrilamid jel elektroforezi (SDS-PAGE),  insan serum PON1’e ait 43 kDa'lık tek bant gösterdi.  Dört antrakinon bileşiği, PON1 aktivitesini, sırasıyla antrorufin, ksantopurpirin, krisazin ve krizofanol için 73, 72.25, 75.19 ve 105.53 M değerleri ile güçlü bir şekilde inhibe etmiştir.   Ki sabitleri sırasıyla 34.64, 39.51, 39.73 ve 63.59 M olarak bulunmuştur.   Tüm antrakinonlar yarışmalı bir şekilde inhibisyon göstermiştir.

Keywords

• Paraoksonaz,inhibisyon,antrokinonlar

Investigation effects of some anthraquinones on human paraoxonase 1 (hPON1)

Abstract

The in vitro effects of the some dihydroxyanthraquinone derivates, anthrorufin, xanthopurpurin, chrysazin and chrysaphanol which are commonly used as anticancer antibiotics, on the activity of purified  serum paraoxonase 1 (PON1) (EC 3.1.8.1.) were investigated.  Paraoxonase 1 was purified from human blood serum 355,12 fold with 55 % yield with use of ammonium sulphate precipitation and Sepharose-4B-L-tyrozine-1-napthylamine HIC (hydrophobic interaction chromatography) coloumn in return.  Sodium Dodecyl Sulphate -polyacrylamide gel electrophoresis (SDS-PAGE) analysis of purified enzyme showed a single band belong to human serum PON1, 43 kDa.  The four anthraquinones inhibited PON activity strongly, with IC50 values for anthrorufin, xanthopurpurin, chrysazin and chrysaphanol of 73, 72.25, 75.19 and 105.53 M respectively.  Ki constants were determined 34.64, 39.51, 39.73 and 63.59 M, respectively.  All anthraquinones showed inhibition in competitive manner.

Keywords

• Paraoxonase,inhibition,anthraquinones

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