"R01 AI110925"
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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.
- 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