A metabolic alkene reporter for spatiotemporally controlled imaging of newly synthesized proteins in Mammalian cells.
The nonsymmetrical spatial distribution of newly synthesized proteins in animal cells plays a central role in many cellular processes. Here, we report that a simple alkene tag, homoallylglycine (HAG), was co-translationally incorporated into a recombinant protein as well as endogenous, newly synthesized proteins in mammalian cells with high efficiency. In conjunction with a photoinduced tetrazole-alkene cycloaddition reaction ("photoclick chemistry"), this alkene tag further served as a bioorthogonal chemical reporter both for the selective protein functionalization in vitro and for a spatiotemporally controlled imaging of the newly synthesized proteins in live mammalian cells. This two-step metabolic alkene tagging-photocontrolled chemical functionalization approach may offer a potentially useful tool to study the role of spatiotemporally regulated protein synthesis in mammalian cells.
Song
Wenjiao
W
Department of Chemistry, State University of New York at Buffalo, 14260, USA.
Wang
Yizhong
Y
Yu
Zhipeng
Z
Vera
Claudia I Rivera
CI
Qu
Jun
J
Lin
Qing
Q
D000501:Allylglycine / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D005434:Flow Cytometry; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D011506:Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*
D011506:Proteins; D000501:Allylglycine