Metabolic syndrome diminishes insulin-induced Akt activation and causes a redistribution of Akt-interacting proteins in cardiomyocytes.

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Insulin signaling in the heart leads to Akt (also known as PKB) activation, a serine/threonine protein kinase, which regulates cardiac glucose metabolism and growth. Cardiac metabolic inflexibility, characterized by impaired insulin-induced glucose uptake and oxidation, has been reported as an early and consistent change in the heart of different models of MetS and diabetes; however, the evaluation of Akt activation has yielded variable results. Here we report in cardiomyocytes of MetS rats, diminished insulin-induced glucose uptake and Akt activation, evaluated by its impaired mobilization towards the plasma membrane and phosphorylation, and reflected in a re-distribution of its interacting proteins, assessed by label-free mass spectrometry (data are available via ProteomeXchange with identifier PXD013260). We report 45 proteins with diminished abundance in Akt complex of MetS cardiomyocytes, mainly represented by energy metabolism-related proteins, and also, 31 Akt-interacting proteins with increased abundance, which were mainly related to contraction, endoplasmic reticulum stress, and Akt negative regulation. These results emphasize the relevance of Akt in the regulation of energy metabolism in the heart and highlight Akt-interacting proteins that could be involved in the detrimental effects of MetS in the heart.", "claims": [{"id": "claim-method", "statement": "ANOVA was used for comparisons of more than two groups, and t-test for unpaired data in the case of two-group comparisons.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:31995605", "locator": "sections[13] \"Statistical analysis\" paragraph 2", "quote": "ANOVA was used for comparisons of more than two groups, and t-test for unpaired data in the case of two-group comparisons."}], "state": "complete"}, {"id": "claim-data-access", "statement": "The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium [34] (http://proteomecentral.proteomexchange.org) via the PRIDE [35] partner repository with the dataset identifier PXD013260.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:31995605", "locator": "sections[18] \"Data accessibility\" paragraph 2", "quote": "The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium [34] (http://proteomecentral.proteomexchange.org) via the PRIDE [35] partner repository with the dataset identifier PXD013260."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": null, "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:31995605"], "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": "complete", "blocked_on": null, "description": "The GEO series accession resolves and its sample accessions are listed. 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Here we report in cardiomyocytes of MetS rats, diminished insulin-induced glucose uptake a", "lin-stimulated cardiomyocytes (N = 3 control rats, N = 3 MetS rats), were loaded on a 10% SDS-", "rdiomyocytes (N = 3 control rats, N = 3 MetS rats), were loaded on a 10% SDS-PAGE. In-gel dige"], "order_basis": "paper_text_order:723", "id": "assay:1"}, {"role": "data_generation", "definition_id": "mass_spectrometry", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["interacting proteins, assessed by label-free mass spectrometry (data are available via ProteomeXchange with", "stically significant. Sample preparation and mass spectrometry analysis Protein samples (30 μg each) from C", "evchenko et al. [30] using 20 ng/μl trypsin (mass spectrometry grade from Sigma-Aldrich). Resulting peptide"], "order_basis": "paper_text_order:966", "id": "assay:2"}, {"role": "data_generation", "definition_id": "western_blot", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.93, "evidence": ["se uptake, Akt-caveolin-3 colocalization and Western blot results. 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