title: Proteome labeling and protein identification in specific tissues and at specific developmental stages in an animal.
abstract: Identifying the proteins synthesized at specific times in cells of interest in an animal will facilitate the study of cellular functions and dynamic processes. Here we introduce stochastic orthogonal recoding of translation with chemoselective modification (SORT-M) to address this challenge. SORT-M involves modifying cells to express an orthogonal aminoacyl-tRNA synthetase/tRNA pair to enable the incorporation of chemically modifiable analogs of amino acids at diverse sense codons in cells in rich media. We apply SORT-M to Drosophila melanogaster fed standard food to label and image proteins in specific tissues at precise developmental stages with diverse chemistries, including cyclopropene-tetrazine inverse electron demand Diels-Alder cycloaddition reactions. We also use SORT-M to identify proteins synthesized in germ cells of the fly ovary without dissection. SORT-M will facilitate the definition of proteins synthesized in specific sets of cells to study development, and learning and memory in flies, and may be extended to other animals.
authors.affiliation: 1] Medical Research Council Laboratory of Molecular Biology, Cambridge, England, UK. [2].
authors.forename: Thomas S
authors.initials: TS
authors.lastname: Elliott
authors.affiliation: 1] Medical Research Council Laboratory of Molecular Biology, Cambridge, England, UK. [2].
authors.forename: Fiona M
authors.initials: FM
authors.lastname: Townsley
authors.affiliation: 1] Medical Research Council Laboratory of Molecular Biology, Cambridge, England, UK. [2].
authors.forename: Ambra
authors.initials: A
authors.lastname: Bianco
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Cambridge, England, UK.
authors.forename: Russell J
authors.initials: RJ
authors.lastname: Ernst
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Cambridge, England, UK.
authors.forename: Amit
authors.initials: A
authors.lastname: Sachdeva
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Cambridge, England, UK.
authors.forename: Simon J
authors.initials: SJ
authors.lastname: Elsässer
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Cambridge, England, UK.
authors.forename: Lloyd
authors.initials: L
authors.lastname: Davis
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Cambridge, England, UK.
authors.forename: Kathrin
authors.initials: K
authors.lastname: Lang
authors.affiliation: 1] Medical Research Council Laboratory of Molecular Biology, Cambridge, England, UK. [2] Department of Chemistry, University of Cambridge, Cambridge, England, UK.
authors.forename: Rudolf
authors.initials: R
authors.lastname: Pisa
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Cambridge, England, UK.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Greiss
authors.affiliation: Department of Biochemistry, University of Cambridge, Cambridge, England, UK.
authors.forename: Kathryn S
authors.initials: KS
authors.lastname: Lilley
authors.affiliation: 1] Medical Research Council Laboratory of Molecular Biology, Cambridge, England, UK. [2] Department of Chemistry, University of Cambridge, Cambridge, England, UK.
authors.forename: Jason W
authors.initials: JW
authors.lastname: Chin
mesh_terms: D000596:Amino Acids / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001709:Biotechnology; D019295:Computational Biology; D004331:Drosophila melanogaster; D015180:Electrophoresis, Gel, Two-Dimensional; D004926:Escherichia coli; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D015335:Molecular Probes; D009928:Organ Specificity; D010053:Ovary / Q000737:chemistry / Q000254:growth & development; D011506:Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D020543:Proteome / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D040901:Proteomics / Q000379:methods*
chemical_list: D000596:Amino Acids; D015335:Molecular Probes; D011506:Proteins; D020543:Proteome
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