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. 2021 Feb 23;11:4367. doi: 10.1038/s41598-021-83666-3

Figure 1.

Figure 1

The probe C15AlkOPP allows labeling of prenylated proteins for detection and proteomic identification. (A) Structures of the native isoprenoids FPP and GGPP and the alkyne-modified analogue C15AlkOPP. (B) Scheme for metabolic labeling with C15AlkOPP in cultured cells. Labeled prenylated proteins are detected through click reaction with a fluorophore and subsequent in-gel fluorescence analysis. Prenylated proteins are enriched through conjugation with biotin and subsequent pulldown with avidin beads. Isolated proteins are digested and peptides are labeled with TMT reagent for identification and quantitation through LC–MS/MS analysis. Quantitative proteomic analysis can be performed using MS2-based or multinotch SPS-MS3 approaches. (C) In-gel fluorescence analysis on COS-7 and HeLa cells treated with 10 μM C15AlkOPP. COS-7 cells display superior labeling compared with HeLa cells. The presence of 10 μM lovastatin enhances probe incorporation whether it is retained (10) or removed (10R) from the culture media during metabolic labeling.