화학공학소재연구정보센터
홈 로그인 로그아웃 연락처 사이트맵
Journal of the Korean Industrial and Engineering Chemistry, Vol.15, No.3, 334-340, May, 2004
Export Citation
HPFA를 이용한 HSA에서 Isoflavone과 Perillyl alcohol 이성질체의 단백질 결합력 측정
Protein Binding of Isoflavones and Enantiomers of Perillyl Alcohol in HSA using High-Performance Frontal Analysis
송명석, 노경호†
  • 인하대학교 초정밀생물분리기술센터, 화학공학과, 인천 402-751
Myong-Seok Song, and Kyung Ho Row†
  • Center for Advanced Bioseparation Technology and Dept. of Chem. Eng., Inha University, Nam-ku, Incheon 402-751, Korea
E-mail:
초록
High-performance frontal analysis (HPFA)를 이용하여 daidzein, genisin, genistein 3 종류의 isoflavones과 R-fom, S-form의 perillyl alcohol 광학이성질체에 대하여 human serum albumin (HSA)과의 단백질 결합력을 연구하였다. 컬럼은 Develosil 100-Diol-5 (10 cm 4.6 mm I.D.)를 사용하였다. Sodium phosphate solution (pH 7.4, ionic strength 0.17)을 1 mL/min의 유량으로 사용하였다. 약리성분인 daidzein과 genisin은 700 μL, genistein은 900 μL perilly alcohol 이성질체는 600 μL에서 크로마토그램이 평평하게 용출되는 것을 알 수 있었고 주입부피로 결정하였다. 실험 데이터는 Scatchard 분석하였다. Isoflavones에 대하여 HSA에 대한 결합 매개변수(K)와 결합능력(nK)은 daidzein이 K=1.581(l/mol), nK=0.771(l/mol)이고, genistein은 K=1.082(l/mol), nK=0.321(l/mol)이었으며, genistin은 K=3.533(l/mol), nK=0.701(l/mol)임을 실험적으로 구하였다. 그리고 perillyl alcohol 이성질체에 대해서는 (S)-perillyl alcohol은 K=5.20106(l/mol), nK=1.74104(l/mol)이었고, (R)-perillyl alcohol은 K=3.71106(l/mol), nK=1.51104(l/mol)로 측정하였다.
High-performance frontal analysis (HPFA) was used for protein binding study of isoflavones (daidzein, genisin, and genistein) and enatiomers of perillyl alcohol (S-POH, R-POH) to human serum albumin (HSA). The analysis was performed on a Develosil 100-Diol-5 (10 cm 4.6 mm I.D.) column. Sodium phosphate solution (pH 7.4, ionic strength 0.17) was used as the mobile phase at a flow rate of 1 mL/min. UV wavelength was set at 260 nm. To ensure the drug to be eluted as a trapezoidal peak with a plateau, injection volumes of 700 μL for daidzein and genisin, 900 μL for genistein were chosen, respectively. Experimental data were fitted by Scatchard analysis. In isoflavones, the binding constant (K) and binding affinity (nK) of them to HSA were: K=1.581105(l/mol), nK=0.77104(l/mol) for daidzein, K=1.082105(l/mol), nK=0.32104(l/mol) for genistein, and K=3.533105(l/mol), nK=0.70104(l/mol) for genistin, respectively. Therefore in enantiomers of perillyl alcohol, the binding constant (K) and binding affinity (nK) to HSA were: K=5.20106(l/mol), nK=1.74104(l/mol) for S-POH and K=3.71106(l/mol), nK=1.51104(l/mol) for R-POH, respectively.
Keywords:HPFA;isoflavones;enantiomers of perillyl alcohol;binding parameter Human Serum Albumin
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