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Application of Peak Intensity Analysis to Measurements of Protein Binding to Lipid Vesicles and Erythrocytes Using Fluorescence Correlation Spectroscopy: Dependence on Particle Size.
- Source :
-
The Journal of membrane biology [J Membr Biol] 2017 Feb; Vol. 250 (1), pp. 77-87. Date of Electronic Publication: 2016 Nov 11. - Publication Year :
- 2017
-
Abstract
- 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.
- Subjects :
- Animals
Biotin
Cattle
Liposomes chemistry
Particle Size
Protein Binding
Proteins metabolism
Ribosome Inactivating Proteins, Type 2 chemistry
Ribosome Inactivating Proteins, Type 2 metabolism
Ricin chemistry
Ricin metabolism
Staining and Labeling
Toxins, Biological chemistry
Toxins, Biological metabolism
Erythrocytes metabolism
Lipids chemistry
Proteins chemistry
Spectrometry, Fluorescence
Subjects
Details
- Language :
- English
- ISSN :
- 1432-1424
- Volume :
- 250
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- The Journal of membrane biology
- Publication Type :
- Academic Journal
- Accession number :
- 27837242
- Full Text :
- https://doi.org/10.1007/s00232-016-9938-6