(A) Chemical structure of the bi-functional NAD+ analog 8-Bu(3-yne)T-NAD+ with the clickable analog sensitivity-inducing, alkyne-containing R group highlighted in red.
(B) Schematic illustrating asPARP activity-dependent, click chemistry-mediated covalent attachment of fluorophores, biotin, or agarose resin to 8-Bu(3-yne)T-ADP-ribosylated proteins.
(C) Automodification reactions with wild-type or analog-sensitive PARP-1, PARP-2, and PARP-3 analyzed by Western blotting for ADP-ribose (top) or click chemistry-based in-gel fluorescence (bottom).