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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Brandão, Hugo B. Sangji, Hussain Pandzic, Elvis Bechstedt, Susanne Brouhard, Gary J. Wiseman, Paul W. |
| Description | Country affiliation: Canada Author Affiliation: Brandão HB ( Department of Physics, McGill University, Montréal, Québec H3A 2T8, Canada.); Sangji H ( Department of Physics, McGill University, Montréal, Québec H3A 2T8, Canada.); Pandzic E ( Department of Physics, McGill University, Montréal, Québec H3A 2T8, Canada.); Bechstedt S ( Department of Biology, McGill University, Montréal, Québec H3A 1B1, Canada.); Brouhard GJ ( Department of Biology, McGill University, Montréal, Québec H3A 1B1, Canada.); Wiseman PW ( Department of Physics, McGill University, Montréal, Québec H3A 2T8, Canada) |
| Abstract | Accurate measurements of kinetic rate constants for interacting biomolecules are crucial for understanding the mechanisms underlying intracellular signalling pathways. The magnitude of binding rates plays a very important molecular regulatory role which can lead to very different cellular physiological responses under different conditions. Here, we extend the k-space image correlation spectroscopy (kICS) technique to study the kinetic binding rates of systems wherein: (a) fluorescently labelled, free ligands in solution interact with unlabelled, diffusing receptors in the plasma membrane and (b) systems where labelled, diffusing receptors are allowed to bind/unbind and interconvert between two different diffusing states on the plasma membrane. We develop the necessary mathematical framework for the kICS analysis and demonstrate how to extract the relevant kinetic binding parameters of the underlying molecular system from fluorescence video-microscopy image time-series. Finally, by examining real data for two model experimental systems, we demonstrate how kICS can be a powerful tool to measure molecular transport coefficients and binding kinetics. |
| File Format | HTM / HTML |
| ISSN | 10462023 |
| Issue Number | 2 |
| Volume Number | 66 |
| e-ISSN | 10959130 |
| Journal | Methods |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2014-03-15 |
| Publisher Place | United States |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Life sciences, Biochemistry Molecular Docking Simulation Animals Cos Cells Cercopithecus Aethiops Cholera Toxin Chemistry Fluorescent Dyes Green Fluorescent Proteins Humans Kinetics Ligands Microscopy, Fluorescence Microtubule-associated Proteins Metabolism Microtubules Neuropeptides Spectrometry, Fluorescence Journal Article Research Support, Non-u.s. Gov't |
| Content Type | Text |
| Resource Type | Article |
| Subject | Molecular Biology |
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