Multi-laboratory experiment PME11 for the standardization of phosphoproteome analysis.

Colomé, Núria; Abian, Joaquín; Aloria, Kerman; Arizmendi, Jesús M; Barceló-Batllori, Silvia; Braga-Lagache, Sophie; Burlet-Schiltz, Odile; Carrascal, Montse; Casal, J Ignacio; Chicano-Gálvez, Eduard; Chiva, Cristina; Clemente, Luis Felipe; Elortza, Felix; Estanyol, Josep M; Fernandez-Irigoyen, Joaquín; Fernández-Puente, Patricia; Fidalgo, María José; Froment, Carine; Fuentes, Manuel; Fuentes-Almagro, Carlos; ... (2022). Multi-laboratory experiment PME11 for the standardization of phosphoproteome analysis. Journal of proteomics, 251, p. 104409. Elsevier 10.1016/j.jprot.2021.104409

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Global analysis of protein phosphorylation by mass spectrometry proteomic techniques has emerged in the last decades as a powerful tool in biological and biomedical research. However, there are several factors that make the global study of the phosphoproteome more challenging than measuring non-modified proteins. The low stoichiometry of the phosphorylated species and the need to retrieve residue specific information require particular attention on sample preparation, data acquisition and processing to ensure reproducibility, qualitative and quantitative robustness and ample phosphoproteome coverage in phosphoproteomic workflows. Aiming to investigate the effect of different variables in the performance of proteome wide phosphoprotein analysis protocols, ProteoRed-ISCIII and EuPA launched the Proteomics Multicentric Experiment 11 (PME11). A reference sample consisting of a yeast protein extract spiked in with different amounts of a phosphomix standard (Sigma/Merck) was distributed to 31 laboratories around the globe. Thirty-six datasets from 23 laboratories were analyzed. Our results indicate the suitability of the PME11 reference sample to benchmark and optimize phosphoproteomics strategies, weighing the influence of different factors, as well as to rank intra and inter laboratory performance.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR)
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Unit Childrens Hospital > Protein- und Zellbiologie

UniBE Contributor:

Braga, Sophie Marie-Pierre, Heller, Manfred

Subjects:

600 Technology > 610 Medicine & health

ISSN:

1874-3919

Publisher:

Elsevier

Language:

English

Submitter:

Marla Rittiner

Date Deposited:

24 Jan 2022 09:31

Last Modified:

05 Dec 2022 16:00

Publisher DOI:

10.1016/j.jprot.2021.104409

PubMed ID:

34758407

BORIS DOI:

10.48350/163437

URI:

https://boris.unibe.ch/id/eprint/163437

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