Quantitative proteomic analysis reveals high interference on protein expression of H9c2 cells activated with glucose and cardiotonic steroids

Meneses-Romero, Erika; Hernández-Orihuela, Lorena; Pando-Robles, Victoria; López, Tomas D.; Oses-Prieto, Juan; Burlingame, Alma L.; Batista, Cesar Vicente Ferreira

doi:10.1016/j.jprot.2019.103536

Cited by 4 publications

(3 citation statements)

References 57 publications

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“…We then performed PRM analysis to validate the DEPs identified by quantitative proteomics. 20 We selected 10 proteins for this analysis, and successfully quantitated and confirmed 8 proteins; 4 proteins were downregulated and 4 were upregulated (Table 1). We selected these proteins for their functional significance based on proteome analysis, and included proteins such as RRAS, which are related to tumor growth and invasion.…”

Section: Validation Of Quantitative Proteomics Results Using Prm and Western Blottingmentioning

confidence: 99%

<p>Quantitative Proteomics Analysis Indicates That Upregulation of lncRNA HULC Promotes Pathogenesis of Glioblastoma Cells</p>

Hu¹,

Ye²,

Li³

et al. 2020

OTT

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Purpose: Glioblastoma (GBM) is an aggressive central nervous system (CNS) cancer and a serious threat to human health. The long noncoding RNA (lncRNA) HULC has been implicated in GBM, but the molecular mechanism is uncertain. This study used quantitative proteomic analysis for global identification of HULC-regulated proteins in glioblastoma cells and identification of potential biomarkers. Materials and Methods: qRT-PCR was used to determine the expression of HULC in U87 cells stably transfected with HULC or an empty vector (control). The CCK-8 assay, transwell assay, and wound-scratch assay were used to measure cell proliferation, invasion, and migration. Quantitative proteomics using Tandem Mass Tag (TMT) labeling, highperformance liquid chromatography (HPLC) fractionation, and liquid chromatographymass spectrometry (LC-MS/MS) analysis were used to identify differentially expressed proteins (DEPs). Screened proteins were validated by parallel reaction monitoring (PRM) and Western blotting. Results: Overexpression of HULC led to increased cell proliferation, invasion, and migration. HULC overexpression also led to significant upregulation of 37 proteins and downregulation of 78 proteins. Bioinformatics analysis indicated these proteins had roles in cellular component, biological process, and molecular function. PRM results of 8 of these proteins (PTK2, TNC, ITGAV, LASP1, MAPK14, ITGA1, GNA13, RRAS) were consistent with the LC-MS/MS and Western blotting results. Conclusion:The results of present study suggest that lncRNA HULC promotes GBM cell proliferation, invasion, and migration by regulating RRAS expression, suggesting that RRAS may be a potential biomarker or therapeutic target for this cancer.

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Section: Validation Of Quantitative Proteomics Results Using Prm and Western Blottingmentioning

confidence: 99%

<p>Quantitative Proteomics Analysis Indicates That Upregulation of lncRNA HULC Promotes Pathogenesis of Glioblastoma Cells</p>

Hu¹,

Ye²,

Li³

et al. 2020

OTT

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“…Both supernatants were mixed in a new tube. The tryptic peptides were dried in a vacuum centrifuge to proceed with desalting using ZipTip C18 (Millipore, Dublin, Ireland) following the manufacturer's protocol [58].…”

Section: Preparation Of Sample For Lc-ms/ms Mass Spectrometrymentioning

confidence: 99%

“…The ions with a charge state +2 to +7, found in the highest abundance, were selectively isolated in a cycle lasting 3 s. The isolation window had a width of 1.2 m/z. Subsequently, they were subjected to fragmentation using higher energy C-trap dissociation (HCD) with a normalized collision energy of 32% [58].…”

Section: Lc-ms/ms Mass Spectrometrymentioning

confidence: 99%

Identification, Biochemical Characterization, and In Vivo Detection of a Zn-Metalloprotease with Collagenase Activity from Mannheimia haemolytica A2

Ramírez-Rico,

Martinez-Castillo,

Ruiz-Mazón

et al. 2024

IJMS

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Respiratory diseases in ruminants are a main cause of economic losses to farmers worldwide. Approximately 25% of ruminants experience at least one episode of respiratory disease during the first year of life. Mannheimia haemolytica is the main etiological bacterial agent in the ruminant respiratory disease complex. M. haemolytica can secrete several virulence factors, such as leukotoxin, lipopolysaccharide, and proteases, that can be targeted to treat infections. At present, little information has been reported on the secretion of M. haemolytica A2 proteases and their host protein targets. Here, we obtained evidence that M. haemolytica A2 proteases promote the degradation of hemoglobin, holo-lactoferrin, albumin, and fibrinogen. Additionally, we performed biochemical characterization for a specific 110 kDa Zn-dependent metalloprotease (110-Mh metalloprotease). This metalloprotease was purified through ion exchange chromatography and characterized using denaturing and chaotropic agents and through zymography assays. Furthermore, mass spectrometry identification and 3D modeling were performed. Then, antibodies against the 110 kDa-Mh metalloprotease were produced, which achieved great inhibition of proteolytic activity. Finally, the antibodies were used to perform immunohistochemical tests on postmortem lung samples from sheep with suggestive histology data of pneumonic mannheimiosis. Taken together, our results strongly suggest that the 110-Mh metalloprotease participates as a virulence mechanism that promotes damage to host tissues.

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Water as a reactant in the differential expression of proteins in cancer

Dick¹

2020

Preprint

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The large amounts of proteomic data now available for cancer can used to investigate whether the physicochemical conditions of tumors are reflected in patterns of protein expression and chemical composition. Compositional analysis of more than 250 datasets for differentially expressed proteins compiled from the literature reveals a clear signal of higher stoichiometric hydration state (n H 2 O , derived from the theoretical formation reactions of proteins from particular basis species) in specific cancer types compared to normal tissue; this trend is also evident in pan-cancer transcriptomic and proteomic datasets from The Cancer Genome Atlas and Human Protein Atlas. In marked contrast to cancer, n H 2 O decreases for differentially expressed proteins in hyperosmotic stress (including high glucose) experiments and 3D cell culture compared to monolayer growth. Compositional analysis combined with gene ages (phylostrata) taken from the literature shows higher n H 2 O of human proteins earlier in evolution. Further analyses using amino acid biosynthetic reactions supports the conclusion that a net increase of water going into the reactions of protein synthesis is a biochemical characteristic shared by most cancer types. These findings raise the possibility of a basic physicochemical link between increased water content in tumors and the atavistic or embryonic patterns of gene and protein expression in cancer.

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Quantitative proteomic analysis reveals high interference on protein expression of H9c2 cells activated with glucose and cardiotonic steroids

Cited by 4 publications

References 57 publications

<p>Quantitative Proteomics Analysis Indicates That Upregulation of lncRNA HULC Promotes Pathogenesis of Glioblastoma Cells</p>

<p>Quantitative Proteomics Analysis Indicates That Upregulation of lncRNA HULC Promotes Pathogenesis of Glioblastoma Cells</p>

Identification, Biochemical Characterization, and In Vivo Detection of a Zn-Metalloprotease with Collagenase Activity from Mannheimia haemolytica A2

Water as a reactant in the differential expression of proteins in cancer

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