Interaction between bovine serum albumin and Indo-1 using fluorescence spectroscopic method
Haixin BAI 1Cheng YANG 2Xiurong YANG2
作者信息
- 1. State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022, China;Graduate University of Chinese Academy of Sciences, Beijing 100039, China
- 2. State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022, China
- 折叠
Abstract
This work attempts to calculate the binding-site number using fluorescence spectroscopic method with bovine serum albumin (BSA) and Indo-1 as proteinand ligand models, respectively. The method for calculat-ing the binding-site number in BSA for Indo-1 was developed based on the relationships between changes in Indo-1 fluorescence intensity and the analytical concen-tration of BSA. The interaction between BSA with Indo-1 was investigated comprehensively using fluorescence techniques as well as fluorescence resonance energy transfer, and the thermodynamic parameters were calculated according to the effect of enthalpy on temperature. Three binding sites in BSA for Indo-1 were revealed, and the distances from Trp212 in BSA to the three binding sites were 2.93, 2.57 and 2.40 nm, respectively. It was also proven that Indo-1 embedded into the three hydrophobic cavities of BSA by hydro-phobic association. This paper provides a reference on calculating the binding-site number in proteins for ligands and studying their interactions by fluorescence spectroscopic methods. In fluorescent quenching experi-ments, fluorescence changes were automatically recorded in real time by combining the Microlab 500 Series Dispenser and PTI fluorescence apparatus.
Key words
bovine serum albumin, Indo-1, binding-site number, interaction, fluorescence spectroscopic methods引用本文复制引用
基金项目
国家自然科学基金(20475052)
国家重点基础研究发展计划(973计划)(2001CB5102)
出版年
2008
NETL
NSTL
万方数据