title: Using SILAC proteomics to investigate the effect of the mycotoxin, alternariol, in the human H295R steroidogenesis model.
abstract: The mycotoxin alternariol (AOH) is an important contaminant of fruits and cereal products. The current study sought to address the effect of a non-toxic AOH concentration on the proteome of the steroidogenic H295R cell model. Quantitative proteomics based on stable isotope labeling by amino acids in cell culture (SILAC) coupled to 1D-SDS-PAGE-LC-MS/MS was applied to subcellular-enriched protein samples. Gene ontology (GO) and ingenuity pathway analysis (IPA) were further carried out for functional annotation and identification of protein interaction networks. Furthermore, the effect of AOH on apoptosis and cell cycle distribution was also determined by the use of flow cytometry analysis. This work identified 22 proteins that were regulated significantly. The regulated proteins are those involved in early stages of steroid biosynthesis (SOAT1, NPC1, and ACBD5) and C21-steroid hormone metabolism (CYP21A2 and HSD3B1). In addition, several proteins known to play a role in cellular assembly, organization, protein synthesis, and cell cycle were regulated. These findings provide a new framework for studying the mechanisms by which AOH modulates steroidogenesis in H295R cell model.
authors.affiliation: Norwegian School of Veterinary Science, Oslo, Norway, skalayout@gmail.com.
authors.forename: Shewit
authors.initials: S
authors.lastname: Kalayou
authors.forename: Anne Grethe
authors.initials: AG
authors.lastname: Hamre
authors.forename: Doreen
authors.initials: D
authors.lastname: Ndossi
authors.forename: Lisa
authors.initials: L
authors.lastname: Connolly
authors.forename: Morten
authors.initials: M
authors.lastname: Sørlie
authors.forename: Erik
authors.initials: E
authors.lastname: Ropstad
authors.forename: Steven
authors.initials: S
authors.lastname: Verhaegen
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism; D000302:Adrenal Cortex / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D017209:Apoptosis / Q000187:drug effects; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D005786:Gene Expression Regulation; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D007553:Isotope Labeling; D007783:Lactones / Q000494:pharmacology*; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D053858:Metabolic Networks and Pathways / Q000187:drug effects; D058977:Molecular Sequence Annotation; D009097:Multienzyme Complexes / Q000235:genetics / Q000378:metabolism; D009183:Mycotoxins / Q000494:pharmacology*; D000077296:Niemann-Pick C1 Protein; D011375:Progesterone Reductase / Q000235:genetics / Q000378:metabolism; D025941:Protein Interaction Mapping; D060066:Protein Interaction Maps / Q000187:drug effects* / Q000235:genetics; D020543:Proteome / Q000235:genetics* / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D013255:Steroid 21-Hydroxylase / Q000235:genetics / Q000378:metabolism; D013251:Steroid Isomerases / Q000235:genetics / Q000378:metabolism; D013256:Steroids / Q000096:biosynthesis*; D002785:Sterol O-Acyltransferase / Q000235:genetics / Q000378:metabolism
chemical_list: C021425:3 beta-hydroxysteroid oxidoreductase-delta(5) 3-ketosteroid isomerase; C548478:ACBD5 protein, human; D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D007783:Lactones; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D009097:Multienzyme Complexes; D009183:Mycotoxins; C106881:NPC1 protein, human; D000077296:Niemann-Pick C1 Protein; D020543:Proteome; D013256:Steroids; D011375:Progesterone Reductase; C531510:CYP21A2 protein, human; D013255:Steroid 21-Hydroxylase; D002785:Sterol O-Acyltransferase; C493492:sterol O-acyltransferase 1; D013251:Steroid Isomerases; C005197:alternariol