Figure 2: Site-specific incorporation of DiZHSeC into proteins in E. coli. | Nature Communications

Figure 2: Site-specific incorporation of DiZHSeC into proteins in E. coli.

From: Genetically encoded protein photocrosslinker with a transferable mass spectrometry-identifiable label

Figure 2

(a) Immunoblotting analysis shows similar amber suppression efficiency of the DiZPK-recognizing PylRS mutant for DiZPK and DiZHSeC when they are inserted to N149 position of GFP (GFP-N149TAG). (The representative result from three replicates are shown). (b) The molecular weight of GFP-N149DiZHSeC is measured by ESI-MS as 27944 Da (calculated 27941 Da). (The representative result from two replicates is shown). (c) DiZHSeC and DiZPK show similar photocrosslinking efficiency on the model protein HdeA. E. coli cells expressing the periplasm-residing HdeA-V58DiZPK or HdeA-V58DiZHSeC protein (carrying a C-terminal His tag) were incubated at pH 2.3 for 30 min followed by ultraviolet irradiation at 365 nm. Cell extracts are separated by the SDS–PAGE gel and analysed by anti-His immunoblotting. The HdeA monomer is marked with a black arrow and the crosslinked complexes are marked with a black brace. (The representative result from three replicates is shown).

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