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Mol. Cells 2011; 31(3): 239-246

Published online January 6, 2011

https://doi.org/10.1007/s10059-011-0026-6

© The Korean Society for Molecular and Cellular Biology

Identification of the Target Proteins of Rosiglitazone in 3T3-L1 Adipocytes through Proteomic Analysis of Cytosolic and Secreted Proteins

Hyun-Ho Hwang1,2, Pyong-Gon Moon1,2, Jeong-Eun Lee1,2, Jung-Guk Kim3, Wan Lee4, Sung-Ho Ryu5, and Moon-Chang Baek1,2,*

1Department of Molecular Medicine, Kyungpook National University, Daegu 700-422, Korea, 2Cell and Matrix Biology Research Institute, Kyungpook National University, Daegu 700-422, Korea, 3Department of Internal Medicine, Kyungpook National University Hospital, Daegu, 700-721, Korea, 4College of Medicine, Dongguk University, Gyeongju 780-714, Korea, 5Pohang University of Science and Technology, Pohang 790-784, Korea

Correspondence to : *Correspondence: mcbaek@knu.ac.kr

Received: September 7, 2010; Revised: November 27, 2010; Accepted: December 6, 2010

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 rosglitazone 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 carboxyki-nase could explain the current action mecha-nism 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.

Keywords 3T3-L1 adipocytes, nano-UPLC MS/MS, proteomic analysis, rosiglitazone

Article

Research Article

Mol. Cells 2011; 31(3): 239-246

Published online March 31, 2011 https://doi.org/10.1007/s10059-011-0026-6

Copyright © The Korean Society for Molecular and Cellular Biology.

Identification of the Target Proteins of Rosiglitazone in 3T3-L1 Adipocytes through Proteomic Analysis of Cytosolic and Secreted Proteins

Hyun-Ho Hwang1,2, Pyong-Gon Moon1,2, Jeong-Eun Lee1,2, Jung-Guk Kim3, Wan Lee4, Sung-Ho Ryu5, and Moon-Chang Baek1,2,*

1Department of Molecular Medicine, Kyungpook National University, Daegu 700-422, Korea, 2Cell and Matrix Biology Research Institute, Kyungpook National University, Daegu 700-422, Korea, 3Department of Internal Medicine, Kyungpook National University Hospital, Daegu, 700-721, Korea, 4College of Medicine, Dongguk University, Gyeongju 780-714, Korea, 5Pohang University of Science and Technology, Pohang 790-784, Korea

Correspondence to:*Correspondence: mcbaek@knu.ac.kr

Received: September 7, 2010; Revised: November 27, 2010; Accepted: December 6, 2010

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 rosglitazone 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 carboxyki-nase could explain the current action mecha-nism 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.

Keywords: 3T3-L1 adipocytes, nano-UPLC MS/MS, proteomic analysis, rosiglitazone

Mol. Cells
Feb 28, 2023 Vol.46 No.2, pp. 69~129
COVER PICTURE
The bulk tissue is a heterogeneous mixture of various cell types, which is depicted as a skein of intertwined threads with diverse colors each of which represents a unique cell type. Single-cell omics analysis untangles efficiently the skein according to the color by providing information of molecules at individual cells and interpretation of such information based on different cell types. The molecules that can be profiled at the individual cell by single-cell omics analysis includes DNA (bottom middle), RNA (bottom right), and protein (bottom left). This special issue reviews single-cell technologies and computational methods that have been developed for the single-cell omics analysis and how they have been applied to improve our understanding of the underlying mechanisms of biological and pathological phenomena at the single-cell level.

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