Proteomic analysis of tyrosine phosphorylation during human liver transplantationReport as inadecuate




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Proteome Science

, 5:1

First Online: 02 January 2007Received: 20 September 2006Accepted: 02 January 2007

Abstract

BackgroundIschemia-reperfusion I-R causes a dramatic reprogramming of cell metabolism during liver transplantation and can be linked to an alteration of the phosphorylation level of several cellular proteins. Over the past two decades, it became clear that tyrosine phosphorylation plays a pivotal role in a variety of important signalling pathways and was linked to a wide spectrum of diseases. Functional profiling of the tyrosine phosphoproteome during liver transplantation is therefore of great biological significance and is likely to lead to the identification of novel targets for drug discovery and provide a basis for novel therapeutic strategies.

ResultsUsing liver biopsies collected during the early phases of organ procurement and transplantation, we aimed at characterizing the global patterns of tyrosine phosphorylation during hepatic I-R. A proteomic approach, based on the purification of tyrosine phosphorylated proteins followed by their identification using mass spectrometry, allowed us to identify Nck-1, a SH2-SH3 adaptor, as a potential regulator of I-R injury. Using immunoblot, cell fractionation and immunohistochemistry, we demonstrate that Nck-1 phosphorylation, expression and localization were affected in liver tissue upon I-R. In addition, mass spectrometry identification of Nck-1 binding partners during the course of the transplantation also suggested a dynamic interaction between Nck-1 and actin during I-R.

ConclusionTaken together, our data suggest that Nck-1 may play a role in I-R-induced actin reorganization, which was previously reported to be detrimental for the hepatocytes of the transplanted graft. Nck-1 could therefore represent a target of choice for the design of new organ preservation strategies, which could consequently help to reduce post-reperfusion liver damages and improve transplantation outcomes.

Abbreviations1D SDS-PAGEOne-dimension Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis

ESIElectroSpray Ionization

GSTGlutathione S-Transferase

I-RIschemia-Reperfusion

kDakiloDaltons

LCLiquid Chromatography

qToFquadrupole Time of Flight

MSMass Spectrometry

MS-MStandem Mass Spectrometry

PBSPhosphate Buffered Saline

PYPhospho-tYrosine

SDStandard Deviation

SH2Src Homology 2

SH3Src Homology 3

Electronic supplementary materialThe online version of this article doi:10.1186-1477-5956-5-1 contains supplementary material, which is available to authorized users.

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Author: Anouk Emadali - Peter P Metrakos - Fariba Kalantari - Tarek Boutros - Daniel Boismenu - Eric Chevet

Source: https://link.springer.com/



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