Cell-selective labeling using amino acid precursors for proteomic studies of multicellular environments.

Abstract:

:We report a technique to selectively and continuously label the proteomes of individual cell types in coculture, named cell type-specific labeling using amino acid precursors (CTAP). Through transgenic expression of exogenous amino acid biosynthesis enzymes, vertebrate cells overcome their dependence on supplemented essential amino acids and can be selectively labeled through metabolic incorporation of amino acids produced from heavy isotope-labeled precursors. When testing CTAP in several human and mouse cell lines, we could differentially label the proteomes of distinct cell populations in coculture and determine the relative expression of proteins by quantitative mass spectrometry. In addition, using CTAP we identified the cell of origin of extracellular proteins secreted from cells in coculture. We believe that this method, which allows linking of proteins to their cell source, will be useful in studies of cell-cell communication and potentially for discovery of biomarkers.

journal_name

Nat Methods

journal_title

Nature methods

authors

Gauthier NP,Soufi B,Walkowicz WE,Pedicord VA,Mavrakis KJ,Macek B,Gin DY,Sander C,Miller ML

doi

10.1038/nmeth.2529

subject

Has Abstract

pub_date

2013-08-01 00:00:00

pages

768-73

issue

8

eissn

1548-7091

issn

1548-7105

pii

nmeth.2529

journal_volume

10

pub_type

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