title: Puromycin Analogues Capable of Multiplexed Imaging and Profiling of Protein Synthesis and Dynamics in Live Cells and Neurons.
abstract: Newly synthesized proteins constitute an important subset of the proteome involved in every cellular process, yet existing chemical tools used to study them have major shortcomings. Herein we report a suite of cell-permeable puromycin analogues capable of being metabolically incorporated into newly synthesized proteins in different mammalian cells, including neuronal cells. Subsequent labeling with suitable bioorthogonal reporters, in both fixed and live cells, enabled direct imaging and enrichment of these proteins. By taking advantage of the mutually orthogonal reactivity of these analogues, we showed multiplexed labeling of different protein populations, as well as quantitative measurements of protein dynamics by fluorescence correlation spectroscopy, could be achieved in live-cell environments.
authors.affiliation: Department of Chemistry, National University of Singapore, 3 Science Drive 3, Singapore, 117543, Singapore.|Institute of Bioengineering, Zhejiang University of Technology, Hangzhou, China.
authors.forename: Jingyan
authors.initials: J
authors.lastname: Ge
authors.affiliation: Key Laboratory of Flexible Electronics & Institute of Advanced Materials, Nanjing Tech University, Nanjing, 211816, China.|National Neuroscience Institute, Singapore, 308433, Singapore.
authors.forename: Cheng-Wu
authors.initials: CW
authors.lastname: Zhang
authors.affiliation: Department of Chemistry, National University of Singapore, 3 Science Drive 3, Singapore, 117543, Singapore.
authors.forename: Xue Wen
authors.initials: XW
authors.lastname: Ng
authors.affiliation: Department of Chemistry, National University of Singapore, 3 Science Drive 3, Singapore, 117543, Singapore.
authors.forename: Bo
authors.initials: B
authors.lastname: Peng
authors.affiliation: Department of Chemistry, National University of Singapore, 3 Science Drive 3, Singapore, 117543, Singapore.
authors.forename: Sijun
authors.initials: S
authors.lastname: Pan
authors.affiliation: Department of Chemistry, National University of Singapore, 3 Science Drive 3, Singapore, 117543, Singapore.
authors.forename: Shubo
authors.initials: S
authors.lastname: Du
authors.affiliation: Department of Chemistry, National University of Singapore, 3 Science Drive 3, Singapore, 117543, Singapore.
authors.forename: Danyang
authors.initials: D
authors.lastname: Wang
authors.affiliation: Key Laboratory of Flexible Electronics & Institute of Advanced Materials, Nanjing Tech University, Nanjing, 211816, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Li
authors.affiliation: National Neuroscience Institute, Singapore, 308433, Singapore.
authors.forename: Kah-Leong
authors.initials: KL
authors.lastname: Lim
authors.affiliation: Department of Chemistry, National University of Singapore, 3 Science Drive 3, Singapore, 117543, Singapore.|Department of Biological Sciences, National University of Singapore, Singapore.
authors.forename: Thorsten
authors.initials: T
authors.lastname: Wohland
authors.affiliation: Department of Chemistry, National University of Singapore, 3 Science Drive 3, Singapore, 117543, Singapore. chmyaosq@nus.edu.sg.
authors.forename: Shao Q
authors.initials: SQ
authors.lastname: Yao
mesh_terms: D006367:HeLa Cells; D006801:Humans; D009474:Neurons / Q000166:cytology*; D014176:Protein Biosynthesis; D011691:Puromycin / Q000737:chemistry*
chemical_list: D011691:Puromycin
keywords: bioorthogonality; metabolic incorporation; multiplexed imaging; newly synthesized proteins; puromycin