Identification of different proteins binding to Na, K-ATPase α1 in LPS-induced ARDS cell model by proteomic analysis.

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:35681168", "kind": "science-record", "revision": 4, "content_hash": "sha256:7f6c527a700dc4588186730913ccfc9b3bbdf61936844af6812ab47c8e441512", "created_at": "2026-11-01T18:14:38Z", "updated_at": "2026-11-01T22:06:43Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "Identification of different proteins binding to Na, K-ATPase α1 in LPS-induced ARDS cell model by proteomic analysis.", "doi": "10.1186/s12953-022-00193-3", "pmid": "35681168", "authors": ["Xu-Peng Wen", "Guo Long", "Yue-Zhong Zhang", "He Huang", "Tao-Hua Liu", "Qi-Quan Wan"], "journal": "Proteome science", "year": 2022, "abstract": "BACKGROUND: Acute respiratory distress syndrome (ARDS) is characterized by refractory hypoxemia caused by accumulation of pulmonary fluid, which is related to inflammatory cell infiltration, impaired tight junction of pulmonary epithelium and impaired Na, K-ATPase function, especially Na, K-ATPase α1 subunit. Up until now, the pathogenic mechanism at the level of protein during lipopolysaccharide- (LPS-) induced ARDS remains unclear.\nMETHODS: Using an unbiased, discovery and quantitative proteomic approach, we discovered the differentially expressed proteins binding to Na, K-ATPase α1 between LPS-A549 cells and Control-A549 cells. These Na, K-ATPase α1 interacting proteins were screened by co-immunoprecipitation (Co-IP) technology. Among them, some of the differentially expressed proteins with significant performance were identified and quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Data are available via ProteomeXchange with identifier PXD032209. The protein interaction network was constructed by the related Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Several differentially expressed proteins were validated by Western blot.\nRESULTS: Of identified 1598 proteins, 89 were differentially expressed proteins between LPS-A549 cells and Control-A549 cells. Intriguingly, protein-protein interaction network showed that there were 244 significantly enriched co-expression among 60 proteins in the group control-A549. while the group LPS-A549 showed 43 significant enriched interactions among 29 proteins. The related GO and KEGG analysis found evident phenomena of ubiquitination and deubiquitination, as well as the pathways related to autophagy. Among proteins with rich abundance, there were several intriguing ones, including the deubiquitinase (OTUB1), the tight junction protein zonula occludens-1 (ZO-1), the scaffold protein in CUL4B-RING ubiquitin ligase (CRL4B) complexes (CUL4B) and the autophagy-related protein sequestosome-1 (SQSTM1).\nCONCLUSIONS: In conclusion, our proteomic approach revealed targets related to the occurrence and development of ARDS, being the first study to investigate significant differences in Na, K-ATPase α1 interacting proteins between LPS-induced ARDS cell model and control-A549 cell. These proteins may help the clinical diagnosis and facilitate the personalized treatment of ARDS.", "claims": [{"id": "claim-method", "statement": "The normalized spectral count of protein in purification was performed as previously described [13]. And the difference multiple selection, identification and quantitative results were as follows: FCA value > 1 or FCA value < -1, and the protein with unique peptide number ≥ 2 was defined as significant difference. Statistical significance was set at a P < 0.05 (*), P < 0.01 (**) and P < 0.001 (***).", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:35681168", "locator": "sections[11] \"Statistical analysis\" paragraph 2", "quote": "The normalized spectral count of protein in purification was performed as previously described [13]. And the difference multiple selection, identification and quantitative results were as follows: FCA value > 1 or FCA value < -1, and the protein with unique peptide number ≥ 2 was defined as significant difference. Statistical significance was set at a P < 0.05 (*), P < 0.01 (**) and P < 0.001 (***)."}], "state": "complete"}, {"id": "claim-data-access", "statement": "The datasets supporting the conclusions of this article are included within the article and its additional files.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:35681168", "locator": "sections[29] \"Availability of data and materials\" paragraph 2", "quote": "The datasets supporting the conclusions of this article are included within the article and its additional files."}], "state": "complete"}, {"id": "claim-target-result", "statement": "Of 1,598 identified proteins, 89 were differentially expressed between LPS-A549 and control-A549 cells, with enrichment analyses implicating ubiquitination, deubiquitination, and autophagy-related proteins including OTUB1, ZO-1, CUL4B, and SQSTM1.", "kind": "qualitative", "target": null, "evidence": [{"asset": "source:pmid:35681168", "locator": "sections[0] \"Abstract\" paragraph 3", "quote": "Of identified 1598 proteins, 89 were differentially expressed proteins between LPS-A549 cells and Control-A549 cells."}, {"asset": "source:pmid:35681168", "locator": "sections[0] \"Abstract\" paragraph 3", "quote": "The related GO and KEGG analysis found evident phenomena of ubiquitination and deubiquitination, as well as the pathways related to autophagy."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": "Of 1,598 identified proteins, 89 were differentially expressed between LPS-A549 and control-A549 cells, with enrichment analyses implicating ubiquitination, deubiquitination, and autophagy-related proteins including OTUB1, ZO-1, CUL4B, and SQSTM1.", "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:35681168"], "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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CONCLUSIONS: In conclusion, our proteomic approach revealed targets related to the occ", "d quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Data are available via ProteomeX"], "order_basis": "paper_text_order:53", "id": "assay:1"}, {"role": "data_generation", "definition_id": "cell_culture", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["teins binding to Na, K-ATPase α1 between LPS-A549 cells and Control-A549 cells. These Na, K-AT", "ATPase α1 between LPS-A549 cells and Control-A549 cells. These Na, K-ATPase α1 interacting pro", "ifferentially expressed proteins between LPS-A549 cells and Control-A549 cells. Intriguingly,", "proteins between LPS-A549 cells and Control-A549 cells. Intriguingly, protein-protein interac"], "order_basis": "paper_text_order:677", "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": ["ntially expressed proteins were validated by Western blot. RESULTS: Of identified 1598 proteins, 89 we"], "order_basis": "paper_text_order:1264", "id": "assay:3"}, {"role": "analysis", "definition_id": "network_analysis", "definition_version": "f2132e96ae6c7a4e", "definition_digest": null, "required_operations": ["build_graph", "centrality_scores", "pathway_enrich"], "support_state": "supported", "source_hashes": {"dsl_catalogue": "f2132e96ae6c7a4e"}, "confidence": 0.7, "evidence": ["protein-protein interaction"], "order_basis": "consumes_earlier_output", "id": "assay:4"}]}