Defining the Akt1 interactome data and delineating alterations in its composition as a function of cell cycle progression.

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Its functional diversity results through interactions with other proteins which change with changing context. This study was designed to capture proteins, which interact with Akt1 as the cell cycle progresses from G0 to G1S and then G2 phase. Such an insight might help us understand the role of Akt1 in cell cycle, which as of now is not well explored. Akt1 expressing HEK 293 cells were cultured in light, medium and heavy labeled SILAC media. Normal lysine and arginine were incorporated as light labels; 6 Da (Dalton) heavier isotopes of the same amino acids were used as medium labels; while for heavy labeling the isotopes were 8 and 10 Da heavier. Light labeled cells were arrested in G0 phase while medium and heavy labeled cells were arrested in G2 and G1S phases, respectively. Equal number of cells from each phase was pooled, lysed and subjected to Affinity Purification coupled to Mass Spectroscopy (AP-MS). The obtained Akt1 protein partners were observed to change as the cell cycle progressed from G0 to G1S and then to G2 phase. Additionally, SILAC labeling aided in quantitative estimation of changing association of a number of proteins which were common to two or more phases, with Akt1. Data are available via ProteomeXchange with identifier PXD005557.", "claims": [{"id": "claim-data-access", "statement": "The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE [6] partner repository with the dataset identifier PXD005557.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:28243621", "locator": "sections[10] \"Database search\" paragraph 4", "quote": "The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE [6] partner repository with the dataset identifier PXD005557."}], "state": "complete"}, {"id": "claim-target-result", "statement": "The Akt1 protein partners changed as the cell cycle progressed, with SILAC labeling enabling quantitative estimation of altered associations; no numerical result for this target is reported, and the source data are available through ProteomeXchange under PXD005557.", "kind": "qualitative", "target": null, "evidence": [{"asset": "source:pmid:28243621", "locator": "sections[0] \"Abstract\" paragraph 1", "quote": "The obtained Akt1 protein partners were observed to change as the cell cycle progressed from G0 to G1S and then to G2 phase."}, {"asset": "source:pmid:28243621", "locator": "sections[0] \"Abstract\" paragraph 1", "quote": "Additionally, SILAC labeling aided in quantitative estimation of changing association of a number of proteins which were common to two or more phases, with Akt1."}, {"asset": "source:pmid:28243621", "locator": "sections[0] \"Abstract\" paragraph 1", "quote": "Data are available via ProteomeXchange with identifier PXD005557."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": "The Akt1 protein partners changed as the cell cycle progressed, with SILAC labeling enabling quantitative estimation of altered associations; no numerical result for this target is reported, and the source data are available through ProteomeXchange under PXD005557.", "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:28243621"], "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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