Reference : Novel post-digest isotope coded protein labeling method for phospho- and glycoproteom...
Scientific journals : Article
Human health sciences : Oncology
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/2268/94423
Novel post-digest isotope coded protein labeling method for phospho- and glycoproteome analysis
English
Fleron, Maximilien mailto [Université de Liège - ULg > Département des sciences biomédicales et précliniques > Histologie - Cytologie >]
Greffe, Yannick mailto [Université de Liège - ULg > > > Doct. sc. bioméd. & pharma. (Bologne)]
Musmeci, Davide mailto [Université de Liège - ULg > > > Doct. sc. bioméd. & pharma. (Bologne)]
Massart, Anne-Cécile [Université de Liège - ULg > > Center for Analytical Research and Technology (CART) >]
Hennequière, Vincent mailto [Université de Liège - ULg > Département des sciences biomédicales et précliniques > Biologie générale et cellulaire >]
Mazzucchelli, Gabriel mailto [Université de Liège - ULg > > Center for Analytical Research and Technology (CART) >]
Waltregny, David mailto [Université de Liège - ULg > Département des sciences cliniques > Urologie >]
De Pauw, Marie-Claire mailto [Université de Liège - ULg > Département des sciences biomédicales et précliniques > Histologie - Cytologie >]
De Pauw, Edwin mailto [Université de Liège - ULg > Département de chimie (sciences) > GIGA-R : Laboratoire de spectrométrie de masse (L.S.M.) >]
Castronovo, Vincenzo mailto [Université de Liège - ULg > Département des sciences biomédicales et précliniques > Biologie générale et cellulaire >]
Turtoi, Andrei mailto [Université de Liège - ULg > Département des sciences biomédicales et précliniques > GIGA-R : Labo de recherche sur les métastases >]
10-Sep-2010
Journal of Proteomics
Elsevier
73
10
1986-2005
Yes (verified by ORBi)
International
1874-3919
1876-7737
Amsterdam
The Netherlands
[en] Protein quantification ; Post-digest ICPL ; Phosphoprotein ; Glycoprotein ; Prostate cancer cells
[en] In the field of proteomics there is an apparent lack of reliable methodology for quantification of posttranslational modifications. Present study offers a novel post-digest ICPL quantification strategy directed towards characterization of phosphorylated and glycosylated proteins. The value of the method is demonstrated based on the comparison of two prostate related metastatic cell lines originating from two distinct metastasis sites (PC3 and LNCaP). The method consists of protein digestion, ICPL labeling, mixing of the samples, PTM enrichment and MS-analysis. Phosphorylated peptides were isolated using TiO(2), whereas the enrichment of glycosylated peptides was performed using hydrazide based chemistry. Isolated PTM peptides were analyzed along with non enriched sample using 2D-(SCX-RP)-Nano-HPLC-MS/MS instrumentation. Taken together the novel ICPL labeling method offered a significant improvement of the number of identified (∼600 individual proteins) and quantified proteins (>95%) in comparison to the classical ICPL method. The results were validated using alternative protein quantification strategies as well as label-free MS quantification method. On the biological level, the comparison of PC3 and LNCaP cells has shown specific modulation of proteins implicated in the fundamental process related to metastasis dissemination. Finally, a preliminary study involving clinically relevant autopsy cases reiterated the potential biological value of the discovered proteins.
Giga-Systems Biology and Chemical Biology ; Giga-Cancer
Fonds pour la formation à la Recherche dans l'Industrie et dans l'Agriculture (Communauté française de Belgique) - FRIA
Researchers
http://hdl.handle.net/2268/94423
10.1016/j.jprot.2010.06.003
http://www.sciencedirect.com/science/article/pii/S1874391910001818

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