Floresans Yöntemler ile Bir Sülfa İlacı ve Pepsin Arasındaki Biyofiziksel Etkileşimler Üzerine Çalışmalar

Abstract

Bir sülfa ilacı olan sülfonamid türevi sülfamerazinin (Smz) pepsin enzimi ile biyofiziksel etkileşimi floresans ve UV absorpsiyon spektroskopileri ile incelenmiştir. Smz'nin pepsine bağlanma modunu araştırmak için kararlı hal ve üç boyutlu (3D) floresans yöntemleri kullanılmıştır ve sonuçlar statik sönümün varlığını ortaya çıkarmıştır. Bağlanma kısım sayıları, bağlanma sabitleri ve termodinamik parametreler farklı sıcaklıklarda hesaplanmıştır. Sonuçlar Smz'nin pepsine kendiliğinden bağlandığını ve hidrofobik/elektrostatik etkileşimlerin önemli rol oynadığını belirtmiştir. 3D spektrumlar, bağlanma modunda pepsin yapısındaki konformasyonel değişiklikleri göstermiştir. Işımasız enerji transferi teorisine dayanarak ilaç ile enzim arasındaki bağlanma mesafesi (r) 3,97 nm olarak hesaplanmıştır. Smz’nin belirlenmesi amacıyla, pepsin varlığında Smz için tespit sınırı (LOD) ve miktar tayin sınırı (LOQ) değerleri sırasıyla 6.54x10-6 M ve 2.18x10-5 M olarak hesaplanmıştır.

Keywords

• Pepsin
• sülfa ilaç
• sülfamerazin
• bağlanma modu
• 3D floresans

Studies on Biophysical Interactions Between a Sulfa Drug and Pepsin by Fluorescence Methods

Abstract

The biophysical interaction of a sulfa drug, sulfonamide derivative sulfamerazine (Smz) with pepsin enzyme was examined using fluorescence and UV-vis absorption spectroscopies. Fluorimetric methods (steady-state/three dimentional fluorescence) were used to examine the binding properties of Smz to pepsin, and the results showed the formation of a complex with a static quenching. Thermodynamic parameters, binding and quenching constants were calculated at different temperatures. The results showed a spontaneous binding between pepsin and Smz through electrostatic/hydrophobic interactions. 3D spectra indicated the conformational/micro environmental changes in the structure of enzyme. Based on the FRET, the binding distance (r) for the drug-enzyme pair was obtained as 3.97 nm. To determine Smz, the limit of detection (LOD) and limit of quantification (LOQ) of Smz were calculated as 6.54x10-6 M and 2.18x10-5 M in the presence of pepsin.

Keywords

• Pepsin
• sulfa drug
• sulfamerazine
• binding mode
• 3D fluorescence

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