title: Identification of the target proteins of rosiglitazone in 3T3-L1 adipocytes through proteomic analysis of cytosolic and secreted proteins.
abstract: Rosiglitazone, one of the thiazolidinedione (TZD), is an oral antidiabetic drug that activates a gamma isoform of peroxisome proliferator-activated receptor (PPARγ). To identify target proteins induced by rosiglitazone in adipocytes, we first performed simultaneous in-depth proteomic profiling of cytosolic proteins and secreted proteins (secretome) from 3T3-L1 adipocytes using a label-free quantification method with nano-UPLC MS/MS. In total, we identified 646 proteins from 3T3-L1 adipocytes, of which 172 and 162 proteins were upregulated and downregulated >1.5-fold, respectively, in rosiglitazone-treated cells, as compared to controls. Some differentially expressed proteins in particular, including fatty acid translocase (FAT)/CD36, fatty acid binding protein, lipoprotein lipase, acetyl CoA acyltransferase, carnitine O-palmitoyltransferase 2, sterol carrier protein, adiponectin, and phosphoenolpyruvate carboxykinase could explain the current action mechanism of TZDs. Furthermore, this study is the first to report on two potential target proteins of rosiglitazone, such as adenomatosis polyposis coli 2 (APC2), and eukaryotic translation initiation factor 5A-1 (eIF5A) related to apoptosis and cell division. Our data clearly suggest that in-depth proteomic approaches using cytosolic and secreted proteins are important and necessary for identification of drug targets at the protein level.
authors.affiliation: Department of Molecular Medicine, Kyungpook National University, Daegu, 700-422, Korea.
authors.forename: Hyun-Ho
authors.initials: HH
authors.lastname: Hwang
authors.forename: Pyong-Gon
authors.initials: PG
authors.lastname: Moon
authors.forename: Jeong-Eun
authors.initials: JE
authors.lastname: Lee
authors.forename: Jung-Guk
authors.initials: JG
authors.lastname: Kim
authors.forename: Wan
authors.initials: W
authors.lastname: Lee
authors.forename: Sung-Ho
authors.initials: SH
authors.lastname: Ryu
authors.forename: Moon-Chang
authors.initials: MC
authors.lastname: Baek
mesh_terms: D041721:3T3-L1 Cells; D017667:Adipocytes / Q000187:drug effects* / Q000378:metabolism; D052242:Adiponectin / Q000378:metabolism; D000818:Animals; D003593:Cytoplasm / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000378:metabolism; D003924:Diabetes Mellitus, Type 2 / Q000235:genetics / Q000378:metabolism; D050556:Fatty Acid-Binding Proteins / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D007328:Insulin / Q000494:pharmacology; D051379:Mice; D009765:Obesity / Q000235:genetics / Q000378:metabolism; D010448:Peptide Initiation Factors / Q000378:metabolism; D020543:Proteome / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism; D000077154:Rosiglitazone; D053719:Tandem Mass Spectrometry; D045162:Thiazolidinediones / Q000378:metabolism* / Q000494:pharmacology; D000097574:Eukaryotic Translation Initiation Factor 5A
chemical_list: D052242:Adiponectin; D003598:Cytoskeletal Proteins; D050556:Fatty Acid-Binding Proteins; D007328:Insulin; D010448:Peptide Initiation Factors; D020543:Proteome; D016601:RNA-Binding Proteins; D000077154:Rosiglitazone; D045162:Thiazolidinediones; D000097574:Eukaryotic Translation Initiation Factor 5A; C499919:Adipoq protein, mouse; C493411:Fabp4 protein, mouse; C117846:adenomatous polyposis coli protein 2