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. Author manuscript; available in PMC: 2018 Aug 1.
Published in final edited form as: Trends Biochem Sci. 2017 May 28;42(8):612–624. doi: 10.1016/j.tibs.2017.05.004

Figure 1. Methods for translation measurement based on luminescent labeling of newly produced peptides.

Figure 1

a. SUnSET (surface sensing of translation) harnesses puromycylated secreted proteins localized on the cell surface for detection by anti-puromycin antibody.

b. Combined with cycloheximide, RPM (ribo-puromycylation) traps puromycylated peptides on the ribosome and visualizes the subcellular sites of translation by staining with an anti-puromycin antibody.

c. Alkyne-bearing puromycin[O-propargyl-puromycin (OP-puro)] allows conjugation of a fluorescent dye to puromycylated proteins by CLICK chemistry.

d. FUNCAT (fluorescent non-canonical amino acid tagging) uses bio-orthogonal amino acids containing a CLICK-reactive alkyne or azide, enabling subsequent fluorophore attachment.

e. PLA (proximity ligation assay) detects newly-synthesized molecules of one target protein by the coincidental localization of a protein-specific antibody with a nascent-protein antibody recognizing puromycin or bio-orthogonal amino acids.

f. Gluc (Gussia luciferase) tagging on the protein of interests monitors its synthesis in real-time with subcellular resolution, by Gluc’s rapid maturation and flash kinetics.