HIV protease (PR) is required for proteolytic maturation in the late phase of HIV replication and represents a prime therapeutic target. The regulation and kinetics of viral polyprotein processing and maturation are currently not understood in detail. Here we design, synthesize, validate and apply a potent, photodegradable HIV PR inhibitor to achieve synchronized induction of proteolysis. The compound exhibits subnanomolar inhibition in vitro. Its photolabile moiety is released on light irradiation, reducing the inhibitory potential by 4 orders of magnitude. We determine the structure of the PR-inhibitor complex, analyze its photolytic products, and show that the enzymatic activity of inhibited PR can be fully restored on inhibitor photolysis. We also demonstrate that proteolysis of immature HIV particles produced in the presence of the inhibitor can be rapidly triggered by light enabling thus to analyze the timing, regulation and spatial requirements of viral processing in real time.
- MeSH
- aminokumariny chemická syntéza farmakologie MeSH
- časové faktory MeSH
- fotolýza MeSH
- HEK293 buňky MeSH
- HIV-1 účinky léků fyziologie účinky záření MeSH
- HIV-proteasa chemie metabolismus MeSH
- inhibitory HIV-proteasy chemická syntéza farmakologie MeSH
- karbamáty chemická syntéza farmakologie MeSH
- kinetika MeSH
- lidé MeSH
- molekulární modely MeSH
- proteinové prekurzory antagonisté a inhibitory chemie metabolismus MeSH
- proteolýza účinky léků MeSH
- replikace viru MeSH
- světlo MeSH
- valin analogy a deriváty chemická syntéza farmakologie MeSH
- vazba proteinů MeSH
- vazebná místa MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
