Kinetic Modeling of mant-ATP Turnover to Interpret the Biochemically Defined Myosin Super-Relaxed State Article Swipe
Filip Ježek
,
Sangwoo Han
,
Alison Schroer Vander Roest
,
Daniel Beard
·
YOU?
·
· 2025
· Open Access
·
· DOI: https://doi.org/10.1101/2025.08.06.667936
YOU?
·
· 2025
· Open Access
·
· DOI: https://doi.org/10.1101/2025.08.06.667936
The fluorescent ATP analog mant-ATP has become a valuable tool for quantifying occupancy of the myosin super-relaxed (SRX) state, a biochemically inactive conformation of myosin in striated muscle. Interpretation of mant-ATP fluorescence decay kinetics is confounded by inconsistencies in state definitions and kinetic assumptions. Here, we develop a mass-action kinetic model of myosin cross-bridge cycling and mant-ATP turnover to reconcile these discrepancies and provide a mechanistic framework for interpreting SRX measurements. Our model simulates ATP label-chase experiments and demonstrates that conventional double-exponential fitting methods do not directly quantify SRX occupancy. Instead, we show that slow and fast decay phases of mant-ATP fluorescence arise from label redistribution among kinetically distinct states, not state populations in equilibrium. The model resolves several apparent paradoxes identified in recent studies by reproducing experimental observations without requiring SRX and DRX kinetic isolation or implausible equilibrium constants. Simulations further quantify the impact of experimental factors—such as ADP accumulation, photobleaching, and initial rigor state occupancy—on fluorescence kinetics and SRX estimates. These results support a revised framework for SRX quantification and suggest that label-chase experiments must be interpreted using mechanistic models to accurately assess myosin state distributions and transition kinetics. Author summary
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Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1101/2025.08.06.667936
- https://www.biorxiv.org/content/biorxiv/early/2025/08/08/2025.08.06.667936.full.pdf
- OA Status
- green
- References
- 17
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4413120107
All OpenAlex metadata
Raw OpenAlex JSON
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https://openalex.org/W4413120107Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.1101/2025.08.06.667936Digital Object Identifier
- Title
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Kinetic Modeling of mant-ATP Turnover to Interpret the Biochemically Defined Myosin Super-Relaxed StateWork title
- Type
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articleOpenAlex work type
- Language
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enPrimary language
- Publication year
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2025Year of publication
- Publication date
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2025-08-08Full publication date if available
- Authors
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Filip Ježek, Sangwoo Han, Alison Schroer Vander Roest, Daniel BeardList of authors in order
- Landing page
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https://doi.org/10.1101/2025.08.06.667936Publisher landing page
- PDF URL
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https://www.biorxiv.org/content/biorxiv/early/2025/08/08/2025.08.06.667936.full.pdfDirect link to full text PDF
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YesWhether a free full text is available
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greenOpen access status per OpenAlex
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https://www.biorxiv.org/content/biorxiv/early/2025/08/08/2025.08.06.667936.full.pdfDirect OA link when available
- Concepts
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Myosin, Kinetics, Fluorescence, Chemistry, Fluorescence recovery after photobleaching, Biophysics, Biological system, Biochemistry, Physics, Biology, Quantum mechanics, MembraneTop concepts (fields/topics) attached by OpenAlex
- Cited by
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0Total citation count in OpenAlex
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17Number of works referenced by this work
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10Other works algorithmically related by OpenAlex
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| abstract_inverted_index.that | 80, 94, 174 |
| abstract_inverted_index.tool | 10 |
| abstract_inverted_index.(SRX) | 18 |
| abstract_inverted_index.Here, | 45 |
| abstract_inverted_index.These | 163 |
| abstract_inverted_index.among | 107 |
| abstract_inverted_index.arise | 103 |
| abstract_inverted_index.decay | 33, 98 |
| abstract_inverted_index.label | 105 |
| abstract_inverted_index.model | 51, 73, 117 |
| abstract_inverted_index.rigor | 155 |
| abstract_inverted_index.state | 40, 112, 156, 187 |
| abstract_inverted_index.these | 61 |
| abstract_inverted_index.using | 180 |
| abstract_inverted_index.Author | 192 |
| abstract_inverted_index.analog | 4 |
| abstract_inverted_index.assess | 185 |
| abstract_inverted_index.become | 7 |
| abstract_inverted_index.impact | 145 |
| abstract_inverted_index.models | 182 |
| abstract_inverted_index.myosin | 16, 25, 53, 186 |
| abstract_inverted_index.phases | 99 |
| abstract_inverted_index.recent | 124 |
| abstract_inverted_index.state, | 19 |
| abstract_inverted_index.cycling | 55 |
| abstract_inverted_index.develop | 47 |
| abstract_inverted_index.fitting | 83 |
| abstract_inverted_index.further | 142 |
| abstract_inverted_index.initial | 154 |
| abstract_inverted_index.kinetic | 43, 50, 135 |
| abstract_inverted_index.methods | 84 |
| abstract_inverted_index.muscle. | 28 |
| abstract_inverted_index.provide | 64 |
| abstract_inverted_index.results | 164 |
| abstract_inverted_index.revised | 167 |
| abstract_inverted_index.several | 119 |
| abstract_inverted_index.states, | 110 |
| abstract_inverted_index.studies | 125 |
| abstract_inverted_index.suggest | 173 |
| abstract_inverted_index.summary | 193 |
| abstract_inverted_index.support | 165 |
| abstract_inverted_index.without | 130 |
| abstract_inverted_index.Abstract | 0 |
| abstract_inverted_index.Instead, | 91 |
| abstract_inverted_index.apparent | 120 |
| abstract_inverted_index.directly | 87 |
| abstract_inverted_index.distinct | 109 |
| abstract_inverted_index.inactive | 22 |
| abstract_inverted_index.kinetics | 34, 159 |
| abstract_inverted_index.mant-ATP | 5, 31, 57, 101 |
| abstract_inverted_index.quantify | 88, 143 |
| abstract_inverted_index.resolves | 118 |
| abstract_inverted_index.striated | 27 |
| abstract_inverted_index.turnover | 58 |
| abstract_inverted_index.valuable | 9 |
| abstract_inverted_index.framework | 67, 168 |
| abstract_inverted_index.isolation | 136 |
| abstract_inverted_index.kinetics. | 191 |
| abstract_inverted_index.occupancy | 13 |
| abstract_inverted_index.paradoxes | 121 |
| abstract_inverted_index.reconcile | 60 |
| abstract_inverted_index.requiring | 131 |
| abstract_inverted_index.simulates | 74 |
| abstract_inverted_index.accurately | 184 |
| abstract_inverted_index.confounded | 36 |
| abstract_inverted_index.constants. | 140 |
| abstract_inverted_index.estimates. | 162 |
| abstract_inverted_index.identified | 122 |
| abstract_inverted_index.occupancy. | 90 |
| abstract_inverted_index.transition | 190 |
| abstract_inverted_index.Simulations | 141 |
| abstract_inverted_index.definitions | 41 |
| abstract_inverted_index.equilibrium | 139 |
| abstract_inverted_index.experiments | 77, 176 |
| abstract_inverted_index.fluorescent | 2 |
| abstract_inverted_index.implausible | 138 |
| abstract_inverted_index.interpreted | 179 |
| abstract_inverted_index.kinetically | 108 |
| abstract_inverted_index.label-chase | 76, 175 |
| abstract_inverted_index.mass-action | 49 |
| abstract_inverted_index.mechanistic | 66, 181 |
| abstract_inverted_index.populations | 113 |
| abstract_inverted_index.quantifying | 12 |
| abstract_inverted_index.reproducing | 127 |
| abstract_inverted_index.assumptions. | 44 |
| abstract_inverted_index.conformation | 23 |
| abstract_inverted_index.conventional | 81 |
| abstract_inverted_index.cross-bridge | 54 |
| abstract_inverted_index.demonstrates | 79 |
| abstract_inverted_index.equilibrium. | 115 |
| abstract_inverted_index.experimental | 128, 147 |
| abstract_inverted_index.fluorescence | 32, 102, 158 |
| abstract_inverted_index.interpreting | 69 |
| abstract_inverted_index.observations | 129 |
| abstract_inverted_index.accumulation, | 151 |
| abstract_inverted_index.biochemically | 21 |
| abstract_inverted_index.discrepancies | 62 |
| abstract_inverted_index.distributions | 188 |
| abstract_inverted_index.measurements. | 71 |
| abstract_inverted_index.super-relaxed | 17 |
| abstract_inverted_index.Interpretation | 29 |
| abstract_inverted_index.factors—such | 148 |
| abstract_inverted_index.occupancy—on | 157 |
| abstract_inverted_index.quantification | 171 |
| abstract_inverted_index.redistribution | 106 |
| abstract_inverted_index.inconsistencies | 38 |
| abstract_inverted_index.photobleaching, | 152 |
| abstract_inverted_index.double-exponential | 82 |
| cited_by_percentile_year | |
| corresponding_author_ids | https://openalex.org/A5054763364 |
| countries_distinct_count | 2 |
| institutions_distinct_count | 4 |
| corresponding_institution_ids | https://openalex.org/I27837315 |
| citation_normalized_percentile.value | 0.48263784 |
| citation_normalized_percentile.is_in_top_1_percent | False |
| citation_normalized_percentile.is_in_top_10_percent | False |