Interaction between bovine serum albumin and Indo-1 using fluorescence spectroscopic method

• 来源期刊: 中国高等学校学术文摘·化学
• 关键词: bovine serum albumin Indo-1 binding-site number interaction fluorescence spectroscopic methods
• 年，卷（期）: 2008,（1）
• 页码范围: 105-111
• 页数: 7页
• 语种: 英文
• DOI: 10.1007/s11458-008-0013-4

摘要

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.