Multiple Click-Selective tRNA Synthetases Expand Mammalian Cell-Specific Proteomics

Andrew C. Yang; Haley Du Bois; Niclas Olsson; David Gate; Benoit Lehallier; Daniela Berdnik; Kyle D. Brewer; Carolyn R. Bertozzi; Joshua E. Elias; Tony Wyss-Coray

doi:10.1021/jacs.8b03074

Multiple Click-Selective tRNA Synthetases Expand Mammalian Cell-Specific Proteomics

Andrew C. Yang, Haley Du Bois, Niclas Olsson, David Gate, Benoit Lehallier, Daniela Berdnik, Kyle D. Brewer, Carolyn R. Bertozzi, Joshua E. Elias, Tony Wyss-Coray^*

^*Corresponding author for this work

Neurology

Research output: Contribution to journal › Article › peer-review

22 Scopus citations

Abstract

Bioorthogonal tools enable cell-type-specific proteomics, a prerequisite to understanding biological processes in multicellular organisms. Here we report two engineered aminoacyl-tRNA synthetases for mammalian bioorthogonal labeling: a tyrosyl (ScTyr_Y43G) and a phenylalanyl (MmPhe_T413G) tRNA synthetase that incorporate azide-bearing noncanonical amino acids specifically into the nascent proteomes of host cells. Azide-labeled proteins are chemoselectively tagged via azide-alkyne cycloadditions with fluorophores for imaging or affinity resins for mass spectrometric characterization. Both mutant synthetases label human, hamster, and mouse cell line proteins and selectively activate their azido-bearing amino acids over 10-fold above the canonical. ScTyr_Y43G and MmPhe_T413G label overlapping but distinct proteomes in human cell lines, with broader proteome coverage upon their coexpression. In mice, ScTyr_Y43G and MmPhe_T413G label the melanoma tumor proteome and plasma secretome. This work furnishes new tools for mammalian residue-specific bioorthogonal chemistry, and enables more robust and comprehensive cell-type-specific proteomics in live mammals.

Original language	English (US)
Pages (from-to)	7046-7051
Number of pages	6
Journal	Journal of the American Chemical Society
Volume	140
Issue number	23
DOIs	https://doi.org/10.1021/jacs.8b03074
State	Published - Jun 13 2018

Funding

We are grateful for financial support by National Institutes of Health grants DP1 AG053015 (T.W.-C.) and R37 GM058867 (C.R.B.), the NOMIS Foundation (T.W.-C.), and the Stanford Neurosciences Institute (T.W.-C., C.R.B., J.E.E.). A.C.Y. was supported by the National Science Foundation Graduate Research Fellowship. N.O. was supported by the Knut and Alice Wallenberg Foundation Postdoctoral Fellowship.

ASJC Scopus subject areas

Catalysis
General Chemistry
Biochemistry
Colloid and Surface Chemistry

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abstract = "Bioorthogonal tools enable cell-type-specific proteomics, a prerequisite to understanding biological processes in multicellular organisms. Here we report two engineered aminoacyl-tRNA synthetases for mammalian bioorthogonal labeling: a tyrosyl (ScTyrY43G) and a phenylalanyl (MmPheT413G) tRNA synthetase that incorporate azide-bearing noncanonical amino acids specifically into the nascent proteomes of host cells. Azide-labeled proteins are chemoselectively tagged via azide-alkyne cycloadditions with fluorophores for imaging or affinity resins for mass spectrometric characterization. Both mutant synthetases label human, hamster, and mouse cell line proteins and selectively activate their azido-bearing amino acids over 10-fold above the canonical. ScTyrY43G and MmPheT413G label overlapping but distinct proteomes in human cell lines, with broader proteome coverage upon their coexpression. In mice, ScTyrY43G and MmPheT413G label the melanoma tumor proteome and plasma secretome. This work furnishes new tools for mammalian residue-specific bioorthogonal chemistry, and enables more robust and comprehensive cell-type-specific proteomics in live mammals.",

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AU - Berdnik, Daniela

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Multiple Click-Selective tRNA Synthetases Expand Mammalian Cell-Specific Proteomics

Abstract

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