Janina Sörmann

Acid-sensing ion channel hASIC1a is persistently activated during ischemia and drives neuronal death, yet small-molecule inhibitors lack selectivity or potency and venom peptides are limited by pH-dependent pharmacology and disulfide-bond production. Using RFdiffusion for backbones, ProteinMPNN for sequences and AlphaFold2 for filtering, the authors narrowed about 12,000 backbones and 70,000 sequences to 76 candidates and identified the 44-amino-acid Denasin1 by automated patch clamp. The miniprotein is disordered in solution, folds into a helix-turn-helix upon binding, inhibits hASIC1a with an IC50 of 23 ± 2.5 nM, and reduced infarct volume from 19.5 ± 9% to 8.6 ± 5.4% in a mouse transient middle cerebral artery occlusion model.

Acid-sensing ion channel hASIC1a is persistently activated during ischemia and drives neuronal death, yet small-molecule inhibitors lack selectivity or potency and venom peptides are limited by pH-dependent pharmacology and disulfide-bond production. Using RFdiffusion for backbones, ProteinMPNN for sequences and AlphaFold2 for filtering, the authors narrowed about 12,000 backbones and 70,000 sequences to 76 candidates and identified the 44-amino-acid Denasin1 by automated patch clamp. The miniprotein is disordered in solution, folds into a helix-turn-helix upon binding, inhibits hASIC1a with an IC50 of 23 ± 2.5 nM, and reduced infarct volume from 19.5 ± 9% to 8.6 ± 5.4% in a mouse transient middle cerebral artery occlusion model.