Post-translational modification patterns on β-myosin heavy chain are altered in ischemic and non-ischemic human hearts
- Florida State University, United States
- University of Washington, United States
- The Ohio State University, United States
- Ohio State University, United States
Abstract
Phosphorylation and acetylation of sarcomeric proteins are important for fine-tuning myocardial contractility. Here, we used bottom-up proteomics and label-free quantification to identify novel post-translational modifications (PTMs) on beta-myosin heavy chain (b-MHC) in normal and failing human heart tissues. We report six acetylated lysines and two phosphorylated residues: K34-Ac, K58-Ac, S210-P, K213-Ac, T215-P, K429-Ac, K951-Ac, and K1195-Ac. K951-Ac was significantly reduced in both ischemic and non-ischemic failing hearts compared to non-diseased hearts. Molecular dynamics simulations show that K951-Ac may impact stability of thick filament tail interactions and ultimately myosin head positioning. K58-Ac altered the solvent exposed SH3 domain surface - known for protein-protein interactions - but did not appreciably change motor domain conformation or dynamics under conditions studied. Together, K213-Ac/T215-P altered loop 1's structure and dynamics - known to regulate ADP-release, ATPase activity, and sliding velocity. Our study suggests that β-MHC acetylation levels may be influenced more by the PTM location than the type of heart disease since less protected acetylation sites are reduced in both heart failure groups. Additionally, these PTMs have potential to modulate interactions between β-MHC and other regulatory sarcomeric proteins, ADP-release rate of myosin, flexibility of the S2 region, and cardiac myofilament contractility in normal and heart failure hearts.
Data availability
All data generated or analyzed during this study are included in the manuscript and the supporting files have been provided for Figures 2, 3, 7, 8 , 9 and Supplemental Figure 1, 2, 4, Tables 1, 2, 3 and 4.Mass spec data have been deposited at Dryad under the unique identifier DOI (doi:10.5061/dryad.s4mw6m97g).
-
Data from: Post-translational modification patterns on β-myosin heavy chain are altered in ischemic and non-ischemic human heartsDryad Digital Repository, doi:10.5061/dryad.s4mw6m97g.
Article and author information
Author details
Matthew C Childers
Michelle S Parvatiyar
Funding
American Heart Association (16SDG2912000)
- Michelle S Parvatiyar
Florida State University (46259)
- Michelle S Parvatiyar
National Institutes of Health (HL128683)
- Jose R Pinto
American Heart Association (2021AHAPRE216237)
- Maicon Landim-Vieira
National Science Foundation (ACI-1548562)
- Michael Regnier
National Institutes of Health (T32HL007828)
- Matthew C Childers
National Institutes of Health (P30AR074990)
- Michael Regnier
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Christopher L-H Huang, University of Cambridge, United Kingdom
Ethics
Human subjects: This study was conducted with the highest ethical standards, human heart samples were collected and stored with full consent of parties involved and were provided by the Lifeline of Ohio with coordination from surgeons and transplant coordinators at the Ohio State University Wexner Medical Center. All aspects of this study were approved and conform to the ethical guidelines established by the Institutional Review Board of The Ohio State University under protocol #2012H0197.
Version history
- Received: October 21, 2021
- Preprint posted: November 22, 2021 (view preprint)
- Accepted: May 1, 2022
- Accepted Manuscript published: May 3, 2022 (version 1)
- Version of Record published: May 20, 2022 (version 2)
Copyright
© 2022, Landim-Vieira et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
Metrics
-
- 1,500
- views
-
- 252
- downloads
-
- 10
- citations
Views, downloads and citations are aggregated across all versions of this paper published by eLife.
Download links
A two-part list of links to download the article, or parts of the article, in various formats.
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
Post-translational modification patterns on β-myosin heavy chain are altered in ischemic and non-ischemic human hearts
eLife 11:e74919.
https://doi.org/10.7554/eLife.74919
Further reading
-
- Structural Biology and Molecular Biophysics
Bacteria utilize various strategies to prevent internal dehydration during hypertonic stress. A common approach to countering the effects of the stress is to import compatible solutes such as glycine betaine, leading to simultaneous passive water fluxes following the osmotic gradient. OpuA from Lactococcus lactis is a type I ABC-importer that uses two substrate-binding domains (SBDs) to capture extracellular glycine betaine and deliver the substrate to the transmembrane domains for subsequent transport. OpuA senses osmotic stress via changes in the internal ionic strength and is furthermore regulated by the 2nd messenger cyclic-di-AMP. We now show, by means of solution-based single-molecule FRET and analysis with multi-parameter photon-by-photon hidden Markov modeling, that the SBDs transiently interact in an ionic strength-dependent manner. The smFRET data are in accordance with the apparent cooperativity in transport and supported by new cryo-EM data of OpuA. We propose that the physical interactions between SBDs and cooperativity in substrate delivery are part of the transport mechanism.
-
- Structural Biology and Molecular Biophysics
Regulated hydrolysis of the phosphoinositide phosphatidylinositol(4,5)-bis-phosphate to diacylglycerol and inositol-1,4,5-P3 defines a major eukaryotic pathway for translation of extracellular cues to intracellular signaling circuits. Members of the lipid-activated protein kinase C isoenzyme family (PKCs) play central roles in this signaling circuit. One of the regulatory mechanisms employed to downregulate stimulated PKC activity is via a proteasome-dependent degradation pathway that is potentiated by peptidyl-prolyl isomerase Pin1. Here, we show that contrary to prevailing models, Pin1 does not regulate conventional PKC isoforms α and βII via a canonical cis-trans isomerization of the peptidyl-prolyl bond. Rather, Pin1 acts as a PKC binding partner that controls PKC activity via sequestration of the C-terminal tail of the kinase. The high-resolution structure of full-length Pin1 complexed to the C-terminal tail of PKCβII reveals that a novel bivalent interaction mode underlies the non-catalytic mode of Pin1 action. Specifically, Pin1 adopts a conformation in which it uses the WW and PPIase domains to engage two conserved phosphorylated PKC motifs, the turn motif and hydrophobic motif, respectively. Hydrophobic motif is a non-canonical Pin1-interacting element. The structural information combined with the results of extensive binding studies and experiments in cultured cells suggest that non-catalytic mechanisms represent unappreciated modes of Pin1-mediated regulation of AGC kinases and other key enzymes/substrates.
