Analysis of the self-assembly process of Aspergillus oryzae hydrophobin RolA by Langmuir-Blodgett method

Hydrophobins are small, amphipathic proteins secreted by filamentous fungi. Hydrophobin RolA, which is produced by Aspergillus oryzae, attaches to solid surfaces, recruits the polyesterase CutL1, and consequently promotes hydrolysis of polyesters. Because this interaction requires the N-terminal, positively charged residue of RolA to be exposed on the solid surface, the orientation of RolA on the solid surface is important for recruitment. However, the process by which RolA forms the self-assembled structure at the interface remains unclear. Using the Langmuir-Blodgett technique, we analyzed the process by which RolA forms a self-assembled structure at the air-water interface and observed the structures on the hydrophobic or hydrophilic SiO₂ substrates via atomic force microscopy. We found that RolA formed self-assembled films in two steps during phase transitions. We observed different assembled structures of RolA on hydrophilic and hydrophobic SiO₂ substrates.