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Not all diazirine proteomics tags photoactivate alike
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.
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What three parts make a typical chemical-proteomic probe?
A recognition group, a reactive photocapture group, and a reporter (fluorophore/affinity/alkyne).
Source: Evaluation of fully-functionalized diazirine tags for chemical proteomic applications · DOI · quote verified
What new minimalist tag did they introduce?
A terminal diazirine of 109 Da made in three steps, 35% overall.
Source: Evaluation of fully-functionalized diazirine tags for chemical proteomic applications · DOI · quote verified
What 365 nm half-lives did LD–F and BD–F show?
5.4 and 2.6 min, first-order.
Source: Evaluation of fully-functionalized diazirine tags for chemical proteomic applications · DOI · quote verified
How does aryl diazirine Ar–F photolyze at 365 nm?
~70% consumed in <2 min, then a slow t½ of 29 min (likely remaining diazo).
Source: Evaluation of fully-functionalized diazirine tags for chemical proteomic applications · DOI · quote verified
Why do DF and Tm fail at 365 nm?
Absorbance maxima shifted to 300–310 nm versus ~360 nm for LD/Ar/BD.
Source: Evaluation of fully-functionalized diazirine tags for chemical proteomic applications · DOI · quote verified
Do DF and Tm photolyze at 302 nm?
Yes—first-order t½ 10 min (DF–F) and 5.8 min (Tm–F).
Source: Evaluation of fully-functionalized diazirine tags for chemical proteomic applications · DOI · quote verified
How many confidently labeled peptides for LD–F, BD–F and Tm–F?
139, 154 and 28 (heavy+light modifications).
Source: Evaluation of fully-functionalized diazirine tags for chemical proteomic applications · DOI · quote verified
What fraction of enriched proteins yielded mapped peptides for LD–F vs BD–F?
11.6% and 11.3% on the benzhydrylazetidine series.
Source: Evaluation of fully-functionalized diazirine tags for chemical proteomic applications · DOI · quote verified
Were staurosporine FF probes cell-permeable?
No—they were used in K562 lysates; JQ1 probes worked in live cells.
Source: Evaluation of fully-functionalized diazirine tags for chemical proteomic applications · DOI · quote verified
Which known staurosporine kinases were co-enriched?
PRKACA, CDK2 and RPS6KA1 (reported Kd 19, 7 and 43 nM).
Source: Evaluation of fully-functionalized diazirine tags for chemical proteomic applications · DOI · quote verified
What live-cell photocrosslinking protocol was used for HEK293T?
20 μM probe, 30 min, then UV.
Source: Evaluation of fully-functionalized diazirine tags for chemical proteomic applications · DOI · quote verified