A Modified FCCS Procedure Applied to Ly49A-MHC Class I cis-Interaction Studies in Cell Membranes
| Autor: | Sofia Johansson, Katja Andersson, Johan Strömqvist, Hideki Muto, Lei Xu, Jerker Widengren, Masataka Kinjo, Yu Ohsugi, Petter Höglund |
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| Rok vydání: | 2011 |
| Předmět: |
EXPRESSION
SURFACE Cell Survival Green Fluorescent Proteins Spectroscopy Imaging and Other Techniques Biophysics Plasma protein binding Human leukocyte antigen Biology Major histocompatibility complex VIVO ACTIVATION Diffusion Cell membrane PROTEIN-PROTEIN INTERACTIONS Mice HLA Antigens Cell Line Tumor MHC class I medicine Animals Receptor Alleles RECEPTOR Cell Membrane FLUCTUATIONS Ligand (biochemistry) LIVE CELLS Biofysik Cell biology FLUORESCENCE CORRELATION SPECTROSCOPY CROSS-CORRELATION SPECTROSCOPY Spectrometry Fluorescence medicine.anatomical_structure biology.protein NK Cell Lectin-Like Receptor Subfamily A Intracellular Protein Binding |
| Zdroj: | Biophysical Journal. 101:1257-1269 |
| ISSN: | 0006-3495 |
| DOI: | 10.1016/j.bpj.2011.06.057 |
| Popis: | The activity of natural killer (NK) cells is regulated by a fine-tuned balance between activating and inhibitory receptors. Dual-color fluorescence cross-correlation spectroscopy (FCCS) was used to directly demonstrate a so-called cis-interaction between a member of the inhibitory NK cell receptor family Ly49 (Ly49A), and its ligand, the major histocompatibility complex (MHC) class I, within the plasma membrane of the same cell. By a refined FCCS model, calibrated by positive and negative control experiments on cells from the same lymphoid cell line, concentrations and diffusion coefficients of free and interacting proteins could be determined on a collection of cells. Using the intrinsic intercellular variation of their expression levels for titration, it was found that the fraction of Ly49A receptors bound in cis increase with increasing amounts of MHC class I ligand. This increase shows a tendency to be more abrupt than for a diffusion limited three dimensional bimolecular reaction, which most likely reflects the two-dimensional confinement of the reaction. For the Ly49A- MHC class I interaction it indicates that within a critical concentration range the local concentration level of MHC class I can provide a distinct regulation mechanism of the NK cell activity. QC 20140609 |
| Databáze: | OpenAIRE |
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