Structure and dynamics of endogenous cardiac troponin complex in human heart tissue captured by native nanoproteomics
Open accessPublished version of a preprint
Abstract
Protein complexes are highly dynamic entities that display substantial diversity in their assembly, post-translational modifications, and non-covalent interactions, allowing them to play critical roles in various biological processes. The heterogeneity, dynamic nature, and low abundance of protein complexes in their native states present challenges to study using conventional structural biology techniques. Here we develop a native nanoproteomics strategy for the enrichment and subsequent native top-down mass spectrometry (nTDMS) analysis of endogenous cardiac troponin (cTn) complex directly from human heart tissue. The cTn complex is enriched and purified using peptide-functionalized superparamagnetic nanoparticles under non-denaturing conditions to enable the isotopic resolution of cTn complex, revealing their complex structure and assembly. Moreover, nTDMS elucidates the stoichiometry and composition of the cTn complex, localizes Ca 2+ binding domains, defines cTn-Ca 2+ binding dynamics, and provides high-resolution mapping of the proteoform landscape. This native nanoproteomics strategy opens a paradigm for structural characterization of endogenous native protein complexes.
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- Authors
- Chapman, E. A., Roberts, D. S., Tiambeng, T. N., Andrews, J., Wang, M.-D., Reasoner, E. A., Melby, J. A., Li, B. H., Kim, D., Alpert, A. J., Jin, S., & Ge, Y.
- Affiliations
- University of Wisconsin–Madison (US)
- Corresponding author
- Jin, Song
- Ge, Ying
- ORCID
- Venue
- Nature Communications
- Published
- 2023-12-18
- Volume
- 14
- Issue
- 1
- Pages
- 8400-8400
- ISSN
- 2041-1723 (ISSN-L)
- PMC
- PMC10728164
- OpenAlex
- W4389894262
- Cited by
- 28
- References
- 16
- Citations by year
- 2026: 10 · 2025: 12 · 2024: 6
- FWCI
- 2.58
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Nature Communications (publishedVersion, cc-by)
- Research Square (acceptedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- PubMed Central (submittedVersion, cc-by)
- RePEc: Research Papers in Economics (submittedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- American Heart Association (832615, R01 HL109810)
- National Institutes of Health (1s10od018475-01)
- Centers for Disease Control and Prevention (S10 OD018475)
- National Heart, Lung, and Blood Institute (R01HL109810, T32HL007936)
- National Institute of General Medical Sciences (R01GM125085, R01GM117058, T32GM008505)
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
- Keywords
- native top-down mass spectrometry, protein complex structure, superparamagnetic nanoparticles, peptide-functionalized nanoparticles, calcium binding, troponin C, troponin I, troponin T, proteoforms, human heart tissue, protein complex stoichiometry
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