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BioMed Research International
Volume 2017, Article ID 8216180, 11 pages
https://doi.org/10.1155/2017/8216180
Research Article

Exogenous Expressions of FTO Wild-Type and R316Q Mutant Proteins Caused an Increase in HNRPK Levels in 3T3-L1 Cells as Demonstrated by DIGE Analysis

1Department of Medical Biology, Kocaeli University Medical School, Umuttepe, 41380 Kocaeli, Turkey
2Department of Biomedical Engineering, Technology Faculty, Kocaeli University, Umuttepe, 41380 Kocaeli, Turkey

Correspondence should be addressed to Murat Kasap; moc.liamg@8002pasakm

Received 23 December 2016; Revised 8 March 2017; Accepted 27 March 2017; Published 7 May 2017

Academic Editor: Rita Casadio

Copyright © 2017 Nil Guzel et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Fat mass and obesity-associated protein is an enzyme that oxidatively demethylates DNA. Although there are numerous studies regarding the catalytic function of FTO, the overall existence or absence of FTO on cellular proteome has not been investigated. This study investigated the changes in the soluble proteome of 3T3-L1 cells upon expression of the WT and the mutant (R316Q) FTO proteins. Protein extracts prepared from 3T3-L1 cells expressing either the WT or the mutant FTO proteins were used in DIGE experiments. Analysis of the data revealed the number of spots matched to every member and there were 350 ± 20 spots with 30.5% overall mean coefficient of variation. Eleven regulated protein spots were excised from the gels and identified by MALDI-TOF/TOF. One of the identified proteins was heterogeneous nuclear ribonucleoprotein K, which displayed more than 2.6- and 3.7-fold increases in its abundance in the WT and the mutant FTO expressing cells, respectively. Western blot analysis validated these observations. This is the first study revealing the presence of a parallel increase in expressions of FTO and HNRNPK proteins. This increase may codictate the metabolic changes occurring in the cell and may attribute a significance to HNRNPK in FTO-associated transformations.