Characterization of the cardiac myosin binding protein-C phosphoproteome in healthy and failing human hearts.

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:23619294", "kind": "science-record", "revision": 5, "content_hash": "sha256:d4cbb25e2fff7bd236485f3278e2b3c750d179d9de5abfdd82b9624a4929947c", "created_at": "2026-11-01T15:40:02Z", "updated_at": "2026-11-01T18:46:54Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "Characterization of the cardiac myosin binding protein-C phosphoproteome in healthy and failing human hearts.", "doi": "10.1016/j.yjmcc.2013.04.012", "pmid": "23619294", "authors": ["Viola Kooij", "Ronald J Holewinski", "Anne M Murphy", "Jennifer E Van Eyk"], "journal": "Journal of molecular and cellular cardiology", "year": 2013, "abstract": "INTRODUCTION: Cardiac myosin binding protein-C (cMyBP-C) becomes dephosphorylated in the failing heart and reduced phosphorylation-dependent regulation of cMyBP-C has been implicated in contractile dysfunction. To date, several phosphorylation sites have been identified for human cMyBP-C; however, a comprehensive characterization of the cMyBP-C phosphoproteome is lacking. This study aimed to characterize the cMyBP-C phosphoproteome using two different proteomic-based methods in explanted donor and end-stage failing hearts.\nMETHODS: The first approach used to characterize the cMyBP-C phosphoproteome employed a strong-cation exchange chromatography (SCX) fractionation method (10 pooled samples, technical replicates=4) and the second employed a sodium dodecylsulfate polyacrylamide gel electrophoresis method (n=10; technical replicates=2). Each subsequently underwent titanium dioxide (TiO2) affinity chromatography to enrich for the tryptic phosphopeptides, which were analyzed using an LTQ-Orbitrap mass spectrometer. Moreover, recombinant C0-C2 fragment of mouse cMyBP-C incubated with PKA, PKC, CaMKII and CK2 was analyzed to identify the kinases involved with phosphorylation of cMyBP-C.\nRESULTS: Seventeen phosphorylation sites on cMyBP-C were identified in vivo, with the majority localized in the N-terminal domains C0-C2. The three most abundant phosphorylated sites, Ser284, Ser286 and Thr290, are located in the regulatory M-domain of cMyBP-C. Ser284 showed a significant reduction in phosphorylation in HF.\nCONCLUSION: This study demonstrates that cMyBP-C harbors more phosphorylation sites than previously known, with a total of 17 (9 novel) identified phosphorylation sites in vivo. Most sites were primarily located within the N-terminal side of the protein. The most highly phosphorylated site on cMyBP-C was Ser284 and this site showed decreased phosphorylation in the failing heart, which implicates importance for fine-tuning contractility. To date, the functional importance of Ser286 and Thr290 is unknown. In addition, 16 sites were identified after in vitro kinase incubation. The data have been deposited to the ProteomeXchange with identifier PXD000158.", "claims": [{"id": "claim-target-result", "statement": "The statistical analysis used t-tests for site-specific phosphorylation comparisons, with Ser284 showing a significant reduction in heart failure (p<0.05); the source data were LC-MS/MS spectral counts from TiO2-enriched phosphopeptides after SDS-PAGE fractionation. The headline result was that Ser284 phosphorylation was significantly reduced in failing hearts.", "kind": "qualitative", "target": null, "evidence": [{"asset": "source:pmid:23619294", "locator": "sections[0] \"Abstract\" paragraph 3", "quote": "Ser284 showed a significant reduction in phosphorylation in HF."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "partial", "objective": "The statistical analysis used t-tests for site-specific phosphorylation comparisons, with Ser284 showing a significant reduction in heart failure (p<0.05); the source data were LC-MS/MS spectral counts from TiO2-enriched phosphopeptides after SDS-PAGE fractionation. The headline result was that Ser284 phosphorylation was significantly reduced in failing hearts.", "steps": [{"id": "read_published_estimate", "order": 1, "name": "Read the published estimate from the source", "kind": "other", "state": "complete", "blocked_on": null, "description": "The claim, method and evidence span published with the source article.", "duration_hours": null, "parameters": {}, "inputs": ["source:pmid:23619294"], "outputs": ["the published estimate and its evidence"], "instruments": [], "reagents": []}, {"id": "obtain_source_data", "order": 2, "name": "Obtain the source data from the named repository", "kind": "data-preparation", "state": "blocked", "blocked_on": "data:accession_absent", "description": "The GEO series accession resolves and its sample accessions are listed. 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Figure 1C shows reduced phosphorylation of", "l enriched tryptic peptides were analyzed by LC-MS/MS on an Agilent 1200 nano-LC system in front o", "-fractionated, TiO2 enriched and analyzed by LC-MS/MS on an LTQ-Orbitrap mass spectrometer. All MS"], "order_basis": "paper_text_order:522", "id": "assay:1"}, {"role": "data_generation", "definition_id": "animal_model", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["phorylation sites on cMyBP-C were identified in vivo, with the majority localized in the N-termin", "7 (9 novel) identified phosphorylation sites in vivo. Most sites were primarily located within th", "ter. Moreover, recombinant C0-C2 fragment of mouse cMyBP-C incubated with PKA, PKC, CaMKII and", "tified a total of 17 phosphorylated residues in vivo. The known sites, Ser133, Ser275, Ser284, an"], "order_basis": "paper_text_order:1133", "id": "assay:2"}]}