Photoaffinity labeling of HIV reverse transcriptase: utilization of 2',3'-dideoxyuridylate analog bearing aryl(trifluoromethyl)diazirine moiety as a chain terminator.

In order to develop a photoaffinity labeling reagent for HIV-1 reverse transcriptase, we utilized 2',3'-dideoxy-E-5-[4-[3-(trifluoromethyl)-3H-diazirin-3-yl] styryl]UTP (TDS-ddUTP) for formation of a photolabile primer. This compound was incorporated into the 3'-terminus of the primer by reverse transcriptase activity, and thus the generated photo-reactive primer was able to bind to the enzyme molecule by photo-irradiation.