MassIVE MSV000093633

Partial Public PXD047719

Native Top-Down Mass Spectrometry for Characterizing Sarcomeric Proteins Directly from Cardiac Tissue Lysate

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Description

Native top-down mass spectrometry is a powerful structural biology tool that can localize post-translational modifications, explore ligand-binding interactions, and elucidate the three-dimensional structure of proteins and protein complexes in the gas-phase. Fourier-transform ion cyclotron resonance-MS offers distinct capabilities for nTDMS, owing to its ultra-high resolving power, mass accuracy, and robust fragmentation techniques. Previous nTDMS studies using FTICR have mainly applied to over-expressed recombinant proteins and protein complexes. Here, we report the first nTDMS study that directly analyzes human heart tissue lysate by direct infusion FTICR-MS without prior chromatographic separation strategies. We have achieved comprehensive nTDMS characterization of cardiac contractile proteins that play critical roles in heart contraction and relaxation. Specifically, our results reveal structural insights into ventricular myosin light chain 2, ventricular myosin light chain 1, and alpha-tropomyosin in the sarcomere, the basic contractile unit of cardiac muscle. Furthermore, we localize the calcium binding domain in MLC-2v. In summary, our nTDMS platform extends the application of FTICR-MS to directly characterize the structure, PTMs, and metal-binding of endogenous proteins from heart tissue lysate without prior separation methods. [doi:10.25345/C5TB0Z59K] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: Native top-down ; cardiac ; FTICR ; endogenous ; structural biology

Contact

Principal Investigators:
(in alphabetical order) 		Ying Ge, University of Wisconsin - Madison, United States
Submitting User: eachapman2
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Distinct protein accessions are counted across all files submitted in the "Statistical Analysis of Quantified Analytes" category having a "Protein" column in this dataset.

"N/A" means no results of this type were submitted.
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