Rhomboid proteases play a variety of physiological roles, but rhomboid protease inhibitors have been mostly developed for the E. coli model rhomboid GlpG. In this work, we screened different electrophilic scaffolds against the human mitochondrial rhomboid PARL and found 4-oxo-β-lactams as submicromolar inhibitors. Multifaceted computations suggest explanations for the activity at the molecular scale and provide models of covalently bound complexes. Together with the straightforward synthesis of the 4-oxo-β-lactam scaffold, this may pave the way toward selective, nonpeptidic PARL inhibitors.

• Publikační typ
• časopisecké články MeSH

Rhomboid intramembrane proteases regulate pathophysiological processes, but their targeting in a disease context has never been achieved. We decoded the atypical substrate specificity of malaria rhomboid PfROM4, but found, unexpectedly, that it results from "steric exclusion": PfROM4 and canonical rhomboid proteases cannot cleave each other's substrates due to reciprocal juxtamembrane steric clashes. Instead, we engineered an optimal sequence that enhanced proteolysis >10-fold, and solved high-resolution structures to discover that boronates enhance inhibition >100-fold. A peptide boronate modeled on our "super-substrate" carrying one "steric-excluding" residue inhibited PfROM4 but not human rhomboid proteolysis. We further screened a library to discover an orthogonal alpha-ketoamide that potently inhibited PfROM4 but not human rhomboid proteolysis. Despite the membrane-immersed target and rapid invasion, ultrastructural analysis revealed that single-dosing blood-stage malaria cultures blocked host-cell invasion and cleared parasitemia. These observations establish a strategy for designing parasite-selective rhomboid inhibitors and expose a druggable dependence on rhomboid proteolysis in non-motile parasites.

• Klíčová slova
• Plasmodium, Ras-converting enzyme, Toxoplasma, apicomplexan parasites, malaria, presenilin, regulated intramembrane proteolysis, rhomboid protease, serine protease, site-2 protease,
• MeSH
• amidy chemická syntéza   chemie   farmakologie   MeSH
• antimalarika chemická syntéza   chemie   farmakologie   MeSH
• HEK293 buňky MeSH
• inhibitory proteas chemická syntéza   chemie   farmakologie   MeSH
• kyseliny boronové chemická syntéza   chemie   farmakologie   MeSH
• lidé MeSH
• malárie krev   farmakoterapie   metabolismus   MeSH
• molekulární struktura MeSH
• parazitické testy citlivosti MeSH
• peptidy chemická syntéza   chemie   farmakologie   MeSH
• Plasmodium falciparum účinky léků   metabolismus   MeSH
• proteasy krev   metabolismus   MeSH
• proteolýza účinky léků   MeSH
• protozoální proteiny antagonisté a inhibitory   krev   metabolismus   MeSH
• racionální návrh léčiv * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Názvy látek
• amidy MeSH
• antimalarika MeSH
• inhibitory proteas MeSH
• kyseliny boronové MeSH
• peptidy MeSH
• proteasy MeSH
• protozoální proteiny MeSH
• ROM4 protein, Plasmodium falciparum MeSH Prohlížeč