CAJ | 학술논문

采用荧光光谱、紫外吸收光谱和圆二色谱(CD)研究了聚乙二醇200基硝苯柳胺与钥孔戚血蓝蛋白(KLH)的相互作用.结果表明,聚乙二醇200基硝苯柳胺对KLH的荧光猝灭机制属于静态猝灭;由Lineweaver-Burk方程计算出不同温度下结合常数K,由Van't Hoff方程计算出△H和△S平均值,结合力主要为静电作用力;根据F(o)rster非辐射能量转移机制求得给体与受体间的结合距离r=5.76 nm;同步荧光光谱表明,聚乙二醇200基硝苯柳胺能够被KLH存储和转运,但结合时对蛋白的构象有一定的影响;圆二色光谱的数据表明相互作用后KLH的二级结构发生了改变:KLH的α-螺旋的含量从43.1%下降到37.8%.
채용형광광보、자외흡수광보화원이색보(CD)연구료취을이순200기초분류알여약공척혈람단백(KLH)적상호작용.결과표명,취을이순200기초분류알대KLH적형광졸멸궤제속우정태졸멸;유Lineweaver-Burk방정계산출불동온도하결합상수K,유Van't Hoff방정계산출△H화△S평균치,결합력주요위정전작용력;근거F(o)rster비복사능량전이궤제구득급체여수체간적결합거리r=5.76 nm;동보형광광보표명,취을이순200기초분류알능구피KLH존저화전운,단결합시대단백적구상유일정적영향;원이색광보적수거표명상호작용후KLH적이급결구발생료개변:KLH적α-라선적함량종43.1%하강도37.8%.
The interaction of keyhole limpet hemocyanin (KLH) and PEG200-niphensamide was investigated by fluorescence spectroscopy, ultraviolet absorption spectroscopy and circular dichroism (CD) spectroscopy. It was shown that KLH fluorescence at 340 nm was quenched regularly with the addition of PEG200-niphensamide; the quenching constants were decreased with the temperature increasing; and the quenching was verified as static fluorescence quenching. The binding constants (K) were obtained according to Lineweaver-Burk equation at different temperature. The calculated thermodynamic parameters (△H, △S) according to Van't Hoff equation indicated that the electrostatic interaction played major roles in the binding processes. The distance between KLH and PEG200-niphensamide was 5.76 nm according to the theory of the F(o)rster energy transference. The effects of PEG200-niphensamide on the conformation of KLH were further analyzed by synchronous fluorescence spectroscopy and circular dichroism spectroscopy. PEG200-niphensamide could be deposited and transported by KLH and it affected the conformation of KLH. The secondary structure of KLH was altered (CD data), I.e., the α-helices were reduced from 43.1% to 37.8%.