Video Sharing

ProteinMPNN can accurately reshape natural proteins, improve solubility, thermal stability, and functional stability while ensuring the protein concept remains unchanged.

Optimization process for stability of human myoglobin nMb:

1. The research team designed and modified the crystal structure of human myoglobin nMb;

2. Firstly, fix the 17 amino acid positions around the heme ligand in the heme binding structure and select the loop region with poor conservation;

3. Use ProteinMPNN for sequence design, modify and reshape the entire protein;

4. Through metal ion affinity chromatography (IMAC) and molecular exclusion chromatography (SEC), it was demonstrated that the total soluble protein yield of most variants increased by 4.1 times;

5. Chromatographic testing shows that these highly expressed variants are structurally identical to natural myoglobin and exhibit higher thermal stability;

Taking dnMb19 as an example, this variant can still maintain complete folding at 95 ℃, while natural myoglobin will collapse at 80 ℃;

7. Compared with natural myoglobin, the dnMb19 variant significantly improves the thermal stability of heme binding;

Researchers further determined the crystal structure of dnMb19 and found a high degree of agreement with the design model;