alexa Epac-induced Alterations in the Proteome of Human SH-SY
ISSN: 0974-276X

Journal of Proteomics & Bioinformatics
Open Access

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Research Article

Epac-induced Alterations in the Proteome of Human SH-SY5Y Neuroblastoma Cells

Even Birkeland1,2, Gyrid Nygaard1,2, Eystein Oveland2, Olav Mjaavatten1,2, Marie Ljones1, Stein Ove Doskeland2, Camilla Krakstad2 and Frode Selheim1,2*

1Proteomic Unit at University of Bergen, Bergen, Norway

2Department of Biomedicine, University of Bergen, Bergen, Norway

*Corresponding Author:
Dr. Frode Selheim
Department of Biomedicine
University of Bergen, Jonas Lies vei 91
N-5009 Bergen, Norway,
Tel: +47 55 586091
Fax: +47 5 5586360
E-mail : [email protected]

Received Date: April 27, 2009; Accepted Date: June 06, 2009; Published Date: June 08, 2009

Citation: Birkeland E, Nygaard G, Oveland E, Mjaavatten O, Ljones M, et al. (2009) Epac-induced Alterations in the Proteome of Human SH-SY5Y Neuroblastoma Cells. J Proteomics Bioinform 2: 244-254. doi: 10.4172/jpb.1000083

Copyright: © 2009 Birkeland E, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

 

Abstract

The hitherto proteome of the human neuroblastoma cell line SH-SY5Y consists of 1029 unique proteins. By using subcellular fractionation, SDS PAGE and nanoLC-Q-TOF-MS/MS we enlarged the proteome with additionally 72.2%, resulting in 3707 unique proteins.

The stage of neuroblastoma tumor cell differentiation is known to influence patient outcome, with a high differentiation stage correlating to favourable prognosis. To elucidate the effects of cAMP in SH-SY5Y neuroblastoma differentiation we used selective cAMP analogs to activate Epac and PKA.

We found that activation of Epac induced actin and tubulin polymerization and neurite outgrowth, whereas PKA activation did not. The effects of the Epac stimulation were abolished by knock down of Epac1 with ShRNA.

Stable isotope labelling with amino acids in cell culture (SILAC) and mass spectrometry were used to disclose the long-time effects of Epac activation on the SH-SY5Y proteome. We found 101 expressed proteins upregulated and 74 downregulated. Upregulated proteins were in general associated with neuronal cell differentiation and adhesion, whereas downregulated proteins typically were involved in RNA processing. We conclude that cAMP-induced morphological and biochemical neuronal differentiation of human neuroblastoma SH-SY5Y cells are mediated by Epac.

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