Week 5 HW: Protein Design Part II

Part A: SOD1 Binder Peptide Design

PeptidePerplexityipTM scoreN terminusB-barrelDimer interface
WRYPAAAAALKX4.308080.3CloseNoSurface bound
WRYGATVAAHKX5.8119530.48FarNoPartially buried
WLSGAAALALKX5.7161310.45CloseNoSurface bound
WLYPAAALALKX8.301710.36FarNoPartially buried
FLYRWLPSRRGG0.38FarNoSurface bound

The predicted protein–peptide complexes produced relatively low ipTM scores overall, indicating weak confidence in the modeled interactions. The PepMLM-generated peptides showed ipTM values ranging from 0.30 to 0.48. The highest score was observed for the peptide WRYGATVAAHKX (ipTM = 0.48), followed by WLSGAAALALKX (ipTM = 0.45), both of which exceeded the ipTM score of the known SOD1-binding peptide FLYRWLPSRRGG (ipTM = 0.38). Despite these slightly higher scores, none of the predicted peptides appeared to strongly interact with the β-barrel region of SOD1, and most were either surface-bound or only partially buried on the protein surface. Overall, while some PepMLM-generated peptides showed marginally higher ipTM scores than the known binder, the predicted interactions remain weak and uncertain.

PeptidePredicted binding affinitySolubilityHemolysis probabilityNet chargeMolecular weight (Da)
WRYPAAAAALKX5.437SolubleNon - hemolitic1.761199.6
WRYGATVAAHKX5.440SolubleNon - hemolitic1.851241.6
WLSGAAALALKX6.550SolubleNon - hemolitic0.761082.6
WLYPAAALALKX6.693SolubleNon - hemolitic0.761198.7
FLYRWLPSRRGG5.96SolubleNon - hemolitic2.761507.7

The peptide property predictions were broadly favorable, since all candidates were predicted to be soluble and non-hemolytic. However, the AlphaFold3 results showed only modest ipTM values, suggesting weak to moderate confidence in the predicted protein-peptide interactions. The peptide with the highest ipTM score was WRYGATVAAHKX (0.48), while the best predicted binding affinity value was observed for WRYPAAAAALKX (5.437), indicating that higher ipTM did not perfectly correlate with stronger predicted affinity. Overall, WRYGATVAAHKX appears to offer the best balance between structural binding potential and therapeutic properties, so it would be the strongest candidate to advance.

I would choose WRYGATVAAHKX, because:

  • it has the highest ipTM
  • it is soluble
  • it is non-hemolytic
  • its charge is moderate
  • it outperformed the known binder in ipTM