MassIVE MSV000090564

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ABRF sPRG 2018 Study: A multipathway phosphopeptide standard for rapid phosphoproteomics assay development

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Description

Recent advances in methodology have made phosphopeptide analysis a tractable problem for many proteomicists. There are now a wide variety of robust and inexpensive enrichment strategies to generate phosphoproteomes, while free or inexpensive software tools for quantitation and site localization have simplified phosphoproteome analysis workflow tremendously. As a research group under the Association for Biomolecular Resource Facilities (ABRF) umbrella, the Proteomics Standards Research Group (sPRG) has worked to develop a multipathway phosphopeptide standard based on a pool of heavy-labeled phosphopeptides designed to enable researchers to rapidly develop assays. This standard contains 131 mass spectrometry vetted phosphopeptides specifically chosen to cover as many known biologically interesting phosphosites as possible from seven different signaling networks: AMPK signaling, death and apoptosis signaling, ErbB signaling, insulin/IGF-1 signaling, mTOR signaling, PI3K/AKT signaling, and stress (p38/SAPK/JNK) signaling. Here we describe a characterization of the standard spiked into a HeLa tryptic digest stimulated with both EGF and IGF1 to activate the MAPK and PI3K/AKT/mTOR pathways. We further demonstrate a comparison of phosphoproteomic profiling of HeLa performed independently in five labs using this standard with data independent acquisition. [doi:10.25345/C54J0B266] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: heavy-labeled phosphopeptides ; phosphoproteomics ; Data independent acquisition ; DIA ; PTM site localization

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Principal Investigators:
(in alphabetical order) 		Brian C. Searle, Ohio State University, USA
Submitting User: sprg
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