Ramazan DEMİRDAĞ, Emrah YERLİKAYA, Ömer İrfan KÜFREVİOĞLU, Cemal GÜNDOĞDU

İnsan Kanserli Kolon Dokusundan CA İzoenzimlerinin Saflaştırılması ve Enzim Aktivitesi Üzerine Bazı Analjezik İlaçların İnhibitör Etkisi

K arbonik anhidraz CA karbon dioksitin karbonik aside hidrasyonundan sorumlu bir enzimdir ve aynı zamanda canlı organizmalarda birçok biyolojik proseslerde yer almaktadır. Bu çalışmada, CA izoenzimleri CA II ve CA IX birlikte sepharose 4B-L-tirozin sülfanilamid afinite kromatografisi kullanılarak 106.67 spesifik aktivite ile % 54.86 verimle 78.4 kat saflaştırıldı. SDS-PAGE’de CA II ve CA IX izoenzimlerin molekül ağırlıkları sırası ile 29 kDa ve 56 kDa olarak hesaplandı. Ayrıca saflaştırılan total enzim üzerine bazı analjezik ilaçların inhibisyon etkisi araştırıldı. Deksketoprofen, pethidin, pheniramidol, tramadol’ün IC50 değerleri sırasıyla 0.0077, 0.025, 0.011, ve 0.04 mM olarak bulundu

Anahtar Kelimeler:

Saflaştırma, Kanser, Kolon, Karbonik Anhidraz, İnhibitör

Purification of Ca Isoenzymes from Human Cancerous Colon Tissue and Inhibitory Effects of Some Analgesics on Enzyme Activity

Carbonic Anhydrase CA is an enzyme which is responsible for the hydration of carbon dioxide to carbonic acid and it also takes places in many biological processes in the living organisms. In this study, CA isoenzymes CA II and CA IX together were purified 78.4 fold with a yield of 54.86 and specific activity of 106.67 by using Sepharose 4B-L-tyrosine sulfanilamide affinity chromatography. In SDS-PAGE molecular weights of CA II and CA IX were calculated as 29 kDa and 56 kDa respectively. Besides inhibitory effects of some analgesics on purified total enzyme was investigated. IC50 values were found as 0.0077, 0.025, 0.011 and 0.04 mM for dexketoprofen, pethidine, phenyramidol and tramadol respectively.

Keywords:

purification, Cancer, Colon, Carbonic anhydrase, Inhibitor,

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