Chemical biology
Not all diazirine proteomics tags photoactivate alike
Open access · cc by · source: Europe PMC
Comparing fully functionalized diazirine tags shows dialkyl tags LD/BD photolyze at 365 nm in minutes, while a new tiny terminal tag needs 302 nm light and labels fewer peptides.
Key findings
LD–F and BD–F t½ = 5.4 and 2.6 min at 365 nm; aryl Ar–F has a 29 min slow phase; DF/Tm barely activate at 365 nm (λmax 300–310 nm) but photolyze at 302 nm (t½ 10 and 5.8 min). LD/BD mapped 11.6/11.3% of enriched proteins to modified peptides versus fewer for Tm.
Methodology
Authors synthesised a 109 Da terminal diazirine, measured 365 vs 302 nm photolysis by LC-MS, then profiled tag-only, fragment, staurosporine and JQ1 probes in HEK293T/K562 proteomes including TMT and crosslink-site mapping.
Limitations
The ‘best’ tag still depends on the recognition ligand; St-series probes were not cell-permeable, so live-cell ranking is incomplete.
How this study connects
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