Cell-specific Profiling of Nascent Proteomes Using Orthogonal Enzyme-mediated Puromycin Incorporation

Ruth M. Barrett, Hui Wen Liu, Haihong Jin, Richard Goodman, Michael Cohen

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Translation regulation is a fundamental component of gene expression, allowing cells to respond rapidly to a variety of stimuli in the absence of new transcription. The lack of methods for profiling nascent proteomes in distinct cell populations in heterogeneous tissues has precluded an understanding of translational regulation in physiologically relevant contexts. Here, we describe a chemical genetic method that involves orthogonal enzyme-mediated incorporation of a clickable puromycin analogue into nascent polypeptides. Using this method, we show that we can label newly synthesized proteins in a cell-specific manner in cells grown in culture and in heterogeneous tissues. We also show that we can identify the nascent proteome in genetically targeted cell populations using affinity enrichment and tandem mass spectrometry. Our method has the potential to provide unprecedented insights into cell-specific translational regulation in heterogeneous tissues.

Original languageEnglish (US)
Pages (from-to)1532-1536
Number of pages5
JournalACS Chemical Biology
Volume11
Issue number6
DOIs
StatePublished - Jun 17 2016

Fingerprint

Puromycin
Proteome
Cells
Tissue
Enzymes
Transcription
Cell culture
Gene expression
Mass spectrometry
Labels
Peptides
Tandem Mass Spectrometry
Population
Proteins
Gene Expression

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

Cite this

Cell-specific Profiling of Nascent Proteomes Using Orthogonal Enzyme-mediated Puromycin Incorporation. / Barrett, Ruth M.; Liu, Hui Wen; Jin, Haihong; Goodman, Richard; Cohen, Michael.

In: ACS Chemical Biology, Vol. 11, No. 6, 17.06.2016, p. 1532-1536.

Research output: Contribution to journalArticle

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