Application of Peak Intensity Analysis to Measurements of Protein Binding to Lipid Vesicles and Erythrocytes Using Fluorescence Correlation Spectroscopy: Dependence on Particle Size

详细信息    查看全文

• 作者：Yuri N. Antonenko ; Anna S. Lapashina ; Elena A. Kotova…
• 关键词：Fluorescence correlation spectroscopy (FCS) ; Ricin ; Viscumin ; Streptavidin ; Liposome ; GM1
• 刊名：The Journal of Membrane Biology
• 出版年：2017
• 出版时间：February 2017
• 年：2017
• 卷：250
• 期：1
• 页码：77-87
• 全文大小：
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Biochemistry, general; Human Physiology;
• 出版者：Springer US
• ISSN：1432-1424
• 卷排序：250

文摘

Fluorescence correlation spectroscopy (FCS) is a sensitive analytical tool for investigation of processes accompanied by changes in the mobility of molecules and complexes. In the present work, peak intensity analysis (PIA) in combination with the solution stirring using FCS setup was applied to explore the interaction between fluorescently labeled protein ligands and corresponding receptors located on membranes. In the system composed of biotinylated liposomes and fluorescently labeled streptavidin as a ligand, PIA allowed us to determine the optimum receptor concentration and demonstrate the essential dependence of the binding efficacy on the length of the linker between the biotin group and the polar head group of the lipid. The binding was dependent on the size of liposomes which was varied by lipid extrusion through filters of different pore diameters. The sensitivity of the method was higher with the liposomes of larger sizes. The PIA approach can be applied not only to liposomes but also to relatively large objects, e.g., erythrocytes or Sepharose beads derivatized with lactose as a receptor for the binding of viscumin and ricin.