not-yet-known not-yet-known not-yet-known unknown Unveiling the self-assembly mechanism of amphiphilic peptide is crucial for the development of functional supramolecular biomaterials. The chemical properties of hydrophilic amino acids play an essential role in this process. Our multiscale molecular dynamic (MD) simulations indicated that the hydrophilic residue, threonine (T) was an excellent candidate to balance the hydrophobicity of the peptide, which could enhance the peptide self-assembly performance. In addition, simulations demonstrated that the number of hydrogen bonds in peptide aggregates was irrelevant to the peptides self-assembly. Avoiding hydrophilic side chains from disrupting the hydrogen bond network between the peptide backbones can improve self-assembly stability. Together with the experimental validation, we believe that T is a promising amino acid to balance the hydrophobicity of amphiphilic peptides. This work highlighted the importance of hydrophilic amino acids in peptide self-assembly, which could be further utilized in designing amphiphilic peptides with different functions.