EFFICIENT AND SENSITIVE ANALYSIS OF BIOMOLECULAR INTERACTIONS BETWEEN PALBOCICLIB AND HUMAN SERUM ALBUMIN USING FLUORESCENCE QUENCHING MECHANISM

Abstract

Objective: In this research, the interaction between Palbociclib (PC) and Human Serum Albumin (HSA) was examined using fluorescence spectroscopy, and the quenching mechanism was elucidated. Material and Method: HSA and PC stock solutions were prepared using 0.1 M pH 7.4 phosphate buffer solution and DMSO, respectively. In the protein-drug interaction experiment, the HSA concentration was fixed at 20 µM, and drug concentrations for the PC-HSA system were used between 0 and 40 µM. Result and Discussion: The interaction between PC and HSA was investigated at various temperatures using spectroscopic techniques. To comprehend the interaction process, experiments were carried out at different physiological pH levels and temperatures. Other parameters, including the number of binding sites, binding constants, and quenching constants, were calculated using the proper formulas. The van't Hoff equation was used to determine thermodynamic parameters such as entropy, Gibbs, and enthalpy at different temperatures.

Keywords

• Drug-protein interaction
• fluorescence spectroscopy
• human serum albumin
• palbociclib
• quenching mechanism

PALBOSİKLİB VE İNSAN SERUM ALBÜMİN ARASINDAKI BİYOMOLEKÜLER ETKİLEŞİMLERİN FLORESAN SÖNÜMLEME MEKANİZMASI KULLANILARAK ETKİLİ VE HASSAS BİR ŞEKİLDE ANALİZ EDİLMESİ

Abstract

Amaç: Bu çalışmada, Palbosiklib (PC) ile İnsan Serum Albümini (HSA) arasındaki etkileşim floresan spektroskopisi kullanılarak incelendi ve sönümleme mekanizması aydınlatıldı. Gereç ve Yöntem: HSA ve PC stok çözeltileri sırasıyla 0.1 M pH 7.4 fosfat tampon çözeltisi ve DMSO kullanılarak hazırlandı. Protein-ilaç etkileşimi deneyinde, HSA konsantrasyonu 20 µM'de sabitlendi ve PC-HSA sistemi için ilaç konsantrasyonları 0 ila 40 µM arasında kullanıldı. Sonuç ve Tartışma: PC ve HSA arasındaki etkileşim, spektroskopik teknikler kullanılarak çeşitli sıcaklıklarda incelenmiştir. Etkileşim sürecini anlamak için farklı fizyolojik pH seviyelerinde ve sıcaklıklarda deneyler gerçekleştirilmiştir. Bağlanma bölgesi sayısı, bağlanma sabitleri ve söndürme sabitleri gibi diğer parametreler, uygun formüller kullanılarak bulunmuştur. van't Hoff denklemi, farklı sıcaklıklarda entropi, Gibbs ve entalpi gibi termodinamik parametrelerini belirlemek için kullanıldı.

Keywords

• Floresan spektroskopisi
• ilaç-protein etkileşimi
• insan serum albümini
• palbosiklib
• sönümleme mekanizması

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