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Methods Mol Biol. 2017;1636:219-234. doi: 10.1007/978-1-4939-7154-1_14.

Cell-Specific Labeling for Analyzing Bidirectional Signaling by Mass Spectrometry.

Author information

1
The Institute of Cancer Research, London, SW3 6JB, UK.
2
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
3
Systems Oncology, CRUK Manchester Institute, The University of Manchester, Wilmslow Road, Manchester, M20 4QL, UK. claus.jorgensen@cruk.manchester.ac.uk.

Abstract

Cell-specific proteome labeling enables global proteome-wide analysis of cell signaling in heterotypic co-cultures. Such approaches have provided unique insight in contact-initiated receptor tyrosine kinase signaling, transfer of proteomic material between heterotypic cells, and interactions between normal and oncogenic cells. Here we describe current methods for cell-specific labeling of heterotypic cells with isotopic labeled amino acids (e.g., SILAC and CTAP). We outline the advantages and disadvantages of individual approaches, describe typical experimental scenarios, and discuss where each experimental approach is optimally applied.

KEYWORDS:

Bidirectional signaling; CTAP; Cell-cell signaling; Heterocellular; PTMs; Phosphoproteomics; SILAC; Signal transduction

PMID:
28730482
DOI:
10.1007/978-1-4939-7154-1_14
[Indexed for MEDLINE]

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