Fenolik Bileşiklerin İnsan Serum Paraoksonaz-1 Enzimi Üzerindeki İnhibisyon Etkileri

Öz

Canlı organizmalardaki metabolik süreçler, enzimlerin katalitik aktivitesi ile yakından ilişkilidir. Enzimlerin inhibisyonu veya indüksiyonu toksisiteye ve metabolik etkileşimlere yol açar.

Bu çalışma, PON1-fenolik madde etkileşimlerini inceleyerek büyüyen ilaç tasarım alanına katkıda bulunmayı amaçlamaktadır. Bu amaçla, paraoksonaz-1 enzimi, hızlı ve farklı kromatografik teknikler

kullanılarak taze insan serumundan saflaştırılmıştır. Ek olarak, bazı fenolik bileşiklerin PON1 üzerindeki inhibisyon etkileri araştırılmış ve saflaştırılan enzimin 3945.15 EU mg-1 protein spesifik

aktiviteye ve 43 kDa molekül ağırlığına sahip olduğu bulunmuştur. Bu bileşikler PON1’e karşı güçlü inhibisyon gösterdiler, özellikle homovanilik asit 13.84±0.08 mM’lık bir IC50 değeri ile PON1’e karşı

önemli bir inhibisyon profili sergiledi. Ki sabitleri homovanillik asit için 6.10±0.26 mM ve phloridzin dihidrat için 16.96±0.76 mM idi. Homovanilik asit yarışmalı inhibisyona sahipken, phloridzin dihidrat

PON1’i yarışmasız olarak inhibe etmiştir. Ayrıca, moleküler yerleştirme hesaplamaları Glide XP modu kullanılarak gerçekleştirilmiştir. Homovanilik asitin glide enerjisi -23.95 kcal mol-1 olarak belirlenmiştir.

Anahtar Kelimeler

• Paraoksonaz,HDL,kromatografi,inhibisyon,fenolik madde,moleküler yerleştirme,moleküler yerleştirme

Inhibition Effects of Phenolic Compounds on Human Serum Paraoxonase-1 Enzyme

Öz

Metabolic processes in living organisms are closely related to the catalytic activity of enzymes. Inhibition or induction of enzymes leads to toxicities and metabolic interactions. This study

aims to contribute to the growing drug design field by studying PON1-phenolic compound interactions. For this purpose, the paraoxonase-1 enzyme was purified from fresh human serum by

using rapid and different chromatographic techniques. Additionally, it was investigated the inhibitory effects of some phenolic substances on the PON1 and was found that the purified enzyme had the

molecular weight of 43 kDa and the specific activity of 3945.15 EU mg-1. These compounds showed potent inhibition against PON1, especially homovanillic acid exhibited a significant inhibition profile

against PON1 with an IC50 value of 13.84±0.08 mM. Ki constants were 6.10±0.26 mM for homovanillic acid and 16.96±0.76 mM for phloridzin dihydrate. Homovanillic acid had competitive

inhibition while the phloridzin dihydrate inhibited the PON1 as non-competitive. Also, molecular docking computations were performed by using the Glide XP mode. Glide energy of the homovanillic

acid determined to be -23.95 kcal mol-1.

Anahtar Kelimeler

• Paraoxonase,HDL,chromatography,inhibition,phenolic compound,molecular docking

Kaynakça

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