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Protein Profile analysis of HeLa Nuclear and Cytoplasmic Subcellular Fractions by Two Dimensional Gel Electrophoresis

[ Vol. 15 , Issue. 3 ]

Author(s):

Esteban Velasquez Saldarriaga, Johanna C. Arroyave Ospina*, Johanna Pedroza-Díaz and Fabian M. Cortes-Mancera   Pages 221 - 228 ( 8 )

Abstract:


Background: HeLa cell line is a reference in vitro model widely used to study processes associated with protein interactions, such as cell signaling, cell cycle progress and apoptosis, among others. However, there are limited publications that include HeLa protein profile characterization through 2D-PAGE.

Objective: The aim of the present report was to describe the differences between total protein profile of HeLa cell line and subcellular fractions obtained from cytoplasm and nucleus.

Methods: Total and subcellular protein fractions were obtained through liquid nitrogen lysis and commercial kit, respectively. Extracted protein was analyzed by 2D-PAGE using the ZOOM IPGRunner system (Invitrogen®) and Image Master Platinum 7.0 (GE®) analysis software for proteome analysis. Identity prediction of spots was performed using Molecular weight (Mw) and Isoelectric point (pI) data from significant spots through Hela 2DE database comparison.

Results: When the integrity of proteins was assessed by SDS-PAGE running, degradation of protein bands was not observed in the gels. In cytoplasmic and Nuclear fractions 324 and 284 spots were detected, versus 202 spots detected in total protein. Subcellular protein profiles also had differences in expression levels, improving the sensitivity of 2D-PAGE. Particularly, in gels of subcellular fractions was found a higher abundance of low molecular weight spots (below 35 kDa) compared to total protein profile.

Conclusion: This report is the first description of HeLa cytoplasmic protein profile characterized by 2D-PAGE approach. Furthermore, the present results will be useful for further research involving protein analysis in this cellular model.

Keywords:

HeLa cells, 2D PAGE, subcellular fractions, proteome, nuclear proteins, cytoplasmic proteins.

Affiliation:

Grupo de Investigacion e Innovacion Biomedica (GI2B), Facultad de Ciencias Exactas y Aplicadas, Instituto Tecnologico Metropolitano (ITM), Medellin, Grupo de Investigacion e Innovacion Biomedica (GI2B), Facultad de Ciencias Exactas y Aplicadas, Instituto Tecnologico Metropolitano (ITM), Medellin, Grupo de Investigacion e Innovacion Biomedica (GI2B), Facultad de Ciencias Exactas y Aplicadas, Instituto Tecnologico Metropolitano (ITM), Medellin, Grupo de Investigacion e Innovacion Biomedica (GI2B), Facultad de Ciencias Exactas y Aplicadas, Instituto Tecnologico Metropolitano (ITM), Medellin

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