PXD060451-1

PXD060451 is an original dataset announced via ProteomeXchange.

Title	Enlightening the decay: a combined experimental and computational analysis of mantATP turnover in skinned muscle fibers
Description	Myosin is the primary motor protein in skeletal muscle, responsible for ATP hydrolysis that drives muscle contraction. In addition to force production, myosin consumes ATP during rest in futile cycles, a phenomenon associated with the Super Relaxed State (SRX), distinct from the disordered Relaxed State (DRX). The SRX is typically measured using the mantATP chasing technique, where the decay of a fluorescent ATP analogue is analyzed and fitted with a combination of exponential decay representing the biochemical states. While this method was initially applied to skinned muscle fibers, it has been adapted for simplersoluble myosin preparations, raising concerns about its reliability. Skinned fibers offer the advantage of preserving the native thick filament structure and myosin cooperativity, while mantATP's limited diffusion and non-specific binding pose challenges. In this study, we combine experimental data and in-silico modeling to dissect the contributions of different components in the mantATP chasing signal. We analyze both rabbit psoas skinned fibers and 'ghost' fibers, where myosin is extracted. Our analysis shows that the fast-decaying component of the signal reflects the effects of diffusion and non-specific binding, occurring on a timescale similar to that of DRX. In contrast, SRX nucleotide release is best described by an exponential decay, with the late timepoints primarily reflecting SRX characteristics. Our findings indicate that mantATP chasing reliably estimates SRX amplitude and time constant, as well as DRX time constant, although it may overestimate DRX amplitude in standard analyses
HostingRepository	PRIDE
AnnounceDate	2025-04-30
AnnouncementXML	Submission_2025-04-30_14:24:44.191.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Luisa Marie Schmidt
SpeciesList	scientific name: Oryctolagus cuniculus (Rabbit); NCBI TaxID: 9986;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Astral

Revision	Datetime	Status	ChangeLog Entry
0	2025-02-04 02:14:08	ID requested
⏵ 1	2025-04-30 14:24:44	announced

Dataset with its publication pending

submitter keyword: MYH extraction, ghost fiber,Single cell, muscle

Leonardo Nogara
contact affiliation	Department of Biomedical Sciences, University of Padua, Padua, Italy Veneto Institute of Molecular Medicine (VIMM), Padua, Italy Department of Pharmaceutical and Pharmacological Sciences, University of Padua, Padua, Italy
contact email	leonardo.nogara@unipd.it
lab head
Luisa Marie Schmidt
contact affiliation	Novo Nordisk Foundation Center for Protein Research, University of Copenhagen
contact email	luisa.schmidt@cpr.ku.dk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD060451
     1. Label: PRIDE project
     2. Name: Enlightening the decay: a combined experimental and computational analysis of mantATP turnover in skinned muscle fibers