Pandzic, Elvis and Morkel, Christian A. and Li, Amy and Cooke, Roger and Whan, Renee M. and dos Remedios, Cristobal G. (2020) Nanomolar ATP binding to single myosin cross-bridges in rigor: a molecular approach to studying myosin ATP kinetics using single human cardiomyocytes. Biophysical Reviews. ISSN 1867-2450
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Nanomolar-ATP-binding-to-single-myosin-crossbridges-in-rigor-a-molecular-approach-to-studying-myosin-ATP-kinetics-using-single-human-cardiomyocytesBiophysical-Reviews.pdf Available under License Creative Commons Attribution No Derivatives. Download (4MB) | Preview |
Abstract
Our knowledge in the field of cardiac muscle and associated cardiomyopathies has been evolving incrementally over the past 60 years and all was possible due to the parallel progress in techniques and methods allowing to take a fresh glimpse at an old problem. Here, we describe an exciting tool used to examine the various states of the human cardiac myosin at the single molecule level. By imaging single Alexa647-ATP binding to permeabilised cardiomyocytes using total internal reflection fluorescence (TIRF) microscopy, we are able to acquire large populations of events in a short timeframe (~ 5000 sites in ~ 10 min) and measure each binding event with high spatio-temporal resolution. The applied algorithm decomposes the point pattern of single molecule binding events into individually distinct binding sites that enables us to recover kinetic parameters, such as bound or free time per site. This single molecule binding approach is a useful tool used to examine muscle contractility. Of particular importance is its application to probing the dynamic lifetimes and proportion of myosins in the super-relaxed state in human cardiomyopathies.
| Item Type: | Article |
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| Subjects: | R Medicine > R Medicine (General) |
| Divisions: | Faculty of Medicine, Health and Life Sciences > School of Medicine |
| Depositing User: | Repository Administrator |
| Date Deposited: | 21 Jul 2020 22:26 |
| Last Modified: | 19 Oct 2021 00:06 |
| URI: | http://eprints.victorchang.edu.au/id/eprint/1006 |
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