Dataset Information

Characterization of hepatocytes secretomes after alpha-amanitin exposure by quantitative proteomics

ABSTRACT: In the human body, proteins secreted into peripheral blood vessels are known as the secretome, and they serve as a representation of the physiological or pathological status of cells. The unique response of cells to toxin exposure can be confirmed via secretome analysis, which can be applied to discover toxic mechanisms or exposure markers. The most widely reported amatoxin is alpha-amanitin (-AMA), which inhibits transcription and protein synthesis by directly interacting with RNA polymerase II. However, the secretory proteins released by hepatic failure caused by -AMA are still not fully characterized. In this study, we analyzed secretory proteins in -AMA-induced hepatotoxicity after -AMA treatment to Huh-7 cells using a comparative proteomics technique. In the results, 1,505 and 1,440 were identified and quantified and significantly up- and down-regulated proteins were 4 and 127 proteins, respectively. Then, bioinformatics analysis was performed to get the GO, Interpro and KEGG pathways about significantly down-regulated proteins. Based on bioinformatics results, we picked complement component 3 (C3) as an exposure marker after -AMA-induced hepatotoxicity. Through western blot and C3 ELISA assay, we validated the C3 level was decreased resulting from -AMA. In conclusion, we found the C3 level in secretomes reduced after -AMA-induced hepatotoxicity using comparative proteomics and molecular biology techniques. We expect this research will be helpful to find new toxic mechanisms, therapeutic targets, and exposure markers in -AMA-induced hepatotoxicity.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hepatocyte

SUBMITTER: Doeun Kim

LAB HEAD: Sangkyu Lee

PROVIDER: PXD036476 | Pride | 2024-05-23

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	20220125_Huh7_5uM_aAMA_Secretome_Con_24h_TMT126-127_HCD_2hr30min_2ug_set1_F1.raw	Raw
	20220125_Huh7_5uM_aAMA_Secretome_Con_24h_TMT126-127_HCD_2hr30min_2ug_set1_F2.raw	Raw
	20220125_Huh7_5uM_aAMA_Secretome_Con_24h_TMT126-127_HCD_2hr30min_2ug_set1_F3.raw	Raw
	20220125_Huh7_5uM_aAMA_Secretome_Con_24h_TMT126-127_HCD_2hr30min_2ug_set1_F4.raw	Raw
	20220125_Huh7_5uM_aAMA_Secretome_Con_24h_TMT126-127_HCD_2hr30min_2ug_set1_F5.raw	Raw

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Publications

Characterization of complement C3 as a marker of alpha-amanitin toxicity by comparative secretome profiling.

Kim Doeun D Lee Min Seo MS Sim Hyunchae H Lee Sangkyu S Lee Hye Suk HS

Toxicological research 20221222 2

In the human body, proteins secreted into peripheral blood vessels are known as the secretome, and they represent the physiological or pathological status of cells. The unique response of cells to toxin exposure can be confirmed <i>via</i> secretome analysis, which can be used to discover toxic mechanisms or exposure markers. Alpha-amanitin (α-AMA) is the most widely studied amatoxin and inhibits transcription and protein synthesis by directly interacting with RNA polymerase II. However, secreto ...[more]

PMID: 37008699

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