Click-MS: tagless protein enrichment using bioorthogonal chemistry for quantitative proteomics

A.H. Smits, A. Borrmann, M. Roosjen, J.C.M. Hest, van, Michiel Vermeulen

Research output: Contribution to journalArticleAcademicpeer-review

10 Citations (Scopus)
292 Downloads (Pure)

Abstract

Epitope-tagging is an effective tool to facilitate protein enrichment from crude cell extracts. Traditionally, N- or C-terminal fused tags are employed, which, however, can perturb protein function. Unnatural amino acids (UAAs) harboring small reactive handles can be site-specifically incorporated into proteins, thus serving as a potential alternative for conventional protein tags. Here, we introduce Click-MS, which combines the power of site-specific UAA incorporation, bioorthogonal chemistry, and quantitative mass spectrometry-based proteomics to specifically enrich a single protein of interest from crude mammalian cell extracts. By genetic encoding of p-azido-l-phenylalanine, the protein of interest can be selectively captured using copper-free click chemistry. We use Click-MS to enrich proteins that function in different cellular compartments, and we identify protein–protein interactions, showing the great potential of Click-MS for interaction proteomics workflows.
Original languageEnglish
Article number12
Pages (from-to)3245–3250
Number of pages6
JournalACS Chemical Biology
Volume11
Issue number12
DOIs
Publication statusPublished - 19 Sep 2016

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