Proteomics

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Proteomics of NGF-TrkA signaling dynamics in neuroblastoma cells

ABSTRACT: We used quantitative mass spectrometry-based proteomics to unravel global nerve growth factor (NGF)-induced TrkA signaling dynamics at the interactome, phosphoproteome and proteome level. A tetracycline-inducible system for TrkA expression was generated in the human neuroblastoma cell line, SH-SY5Y. TrkA-induced cells were stimulated with NGF for different time points to follow phosphoproteome, interactome and proteome changes on a temporal scale. In a triple SILAC setup (Light: Lys0,Arg0; Medium: Lys4,Arg6; and Heavy: Lys8,Arg10), the samples were stimulated with NGF as indicated. Phosphoproteome: 0, 10, 45 min and 0, 120, 120 min+cycloheximide. Interactome: 0, 5, 10 min. Proteome: 0, 24, 48 h. All experiments were performed as biological replicates.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Jesper V. Olsen  

PROVIDER: MSV000080834 | MassIVE | Fri Mar 31 17:19:00 BST 2017

SECONDARY ACCESSION(S): PXD001115

REPOSITORIES: MassIVE

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Temporal proteomics of NGF-TrkA signaling identifies an inhibitory role for the E3 ligase Cbl-b in neuroblastoma cell differentiation.

Emdal Kristina B KB   Pedersen Anna-Kathrine AK   Bekker-Jensen Dorte B DB   Tsafou Kalliopi P KP   Horn Heiko H   Lindner Sven S   Schulte Johannes H JH   Eggert Angelika A   Jensen Lars J LJ   Francavilla Chiara C   Olsen Jesper V JV  

Science signaling 20150428 374

SH-SY5Y neuroblastoma cells respond to nerve growth factor (NGF)-mediated activation of the tropomyosin-related kinase A (TrkA) with neurite outgrowth, thereby providing a model to study neuronal differentiation. We performed a time-resolved analysis of NGF-TrkA signaling in neuroblastoma cells using mass spectrometry-based quantitative proteomics. The combination of interactome, phosphoproteome, and proteome data provided temporal insights into the molecular events downstream of NGF binding to  ...[more]

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