İnsan Eritrositlerinden Saflaştırılan Karbonik Anhidraz (CA) I ve II İzoenzimleri Üzerinde Eozin Y'nin İnhibisyon Etkisi

Bütün hücrelerde metabolizma sonucu açığa çıkan ve vücuttan atılması gereken karbondioksit üretilir. Bu CO2'nin büyük bir kısmı eritrositlerde CA enzimi tarafından bikarbonata dönüştürülür ve vücuttan atılır. Bu nedenle, CA'nın kırmızı kan hücrelerinde hayati bir rolü vardır. Ayrıca, CA diğer birçok patolojik ve fizyolojik süreçlere dahil olmuştur ve CA inhibitörlerinin, özellikle glokom gibi birçok hastalığın tedavisinde ve tanısında etkili olduğu belirlenmiştir. CA inhibitörlerinin önemi göz önüne alındığında, bu çalışmada Eosin Y'nin insan eritrositlerinden saflaştırılmış CA I ve CA II izoenzimlerin aktivitesi üzerindeki inhibisyon etkilerinin araştırılması amaçlanmıştır. Eozin Y, histolojik ve tıbbi uygulamalarda yaygın olarak kullanılan bir boya molekülüdür. Bu amaçla öncelikle CA I ve CA II izoenzimleri, Sepharose-4B-L-tirozin-sülfanilamid afinite kromatografısi kullanılarak insan eritrositlerinden saflaştırıldı. Daha sonra, Eozin Y'nin bu insan eritrosit CA I (hCA I) ve CA II (hCA II) izoenzimlerinin aktivitesi üzerindeki inhibitör etkisi araştırıldı. hCA I ve hCA II'nin millimolar aralıktaki Eozin Y tarafından inhibe edildiği belirlendi. IC50 değerleri hCA I için 3,78 mM ve hCA II için 2,04 mM. Ki değerleri ise hCA I ve hCA II için sırasıyla 9,65 ± 0,968 mM ve 7,52 ± 2,88 mM olarak tespit edildi. Sonuç olarak, bu çalışmada elde edilen sonuçların, ilaç adayı olabilecek yeni CA inhibitörlerinin geliştirilmesinde faydalı olacağı ümit edilmektedir.

Anahtar Kelimeler:

İnsan eritrositi, karbonik anhidraz, eozin Y, inhibisyon

Inhibition Effect of Eosin Y on Carbonic Anhydrase (CA) I and II Isoenzymes Purified from Human Erythrocytes

All cells produce carbon dioxide (CO2), which is released as a result of metabolism and must be removed from the body. A large part of this CO2 is converted to bicarbonate by the carbonic anhydrase (CA) enzyme in erythrocytes and is discarded from the body. So, CA has a vital role in red blood cells. In addition to, CA involved in many other pathological and physiological processes and it was determined that the inhibitors of CA were effective in the treatment and diagnosis of many diseases particularly glaucoma. Considering the importance of the CA's inhibitors, in this study it was intended to research the inhibition effects of Eosin Y on CA I and CA II isoenzymes activity purified from human erythrocytes. Eosin Y is a dye molecule commonly used in histological and medical applications. For this purpose, firstly CA I and CA II isoenzymes were purified from human erythrocytes by using Sepharose-4B-L-tyrosine-sulfanilamide affinity chromatography. Then the inhibitory effect of Eosin Y on the activity of these human erythrocyte CA I (hCA I) and CA II (hCA II) isoenzymes was investigated. It was determined that hCA I and hCA II were inhibited by Eosin Y in the millimolar range. IC50 values were found to be 3.78 mM for hCA I and 2.04 mM for hCA II and Ki values were determined as 9.65±0.968 mM and 7.52±2.88 mM for hCA I and hCA II, respectively. In conclusion, it is hoped that the results obtained in this study may be beneficial in the development of new CA inhibitors which may be drug candidates.

Keywords:

Human erythrocyte, carbonic anhydrase, eosin Y, inhibition,

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