Structure Formation in Class†I and Class†II Hydrophobins at the Air-Water Interface under Multiple Compression/Expansion Cycles.

Hydrophobins are small amphiphilic fungal proteins empirically divided into two classes. We investigated the self-assembled structures of class I SC3 from S. commune and class II HFBII from T. reesei transferred to mica from the air-water interface by using the Langmuir-Schaefer (LS) technique and atomic force microscopy (AFM). The main focus is the influence of areal constraint and multiple compressions and expansions on the morphology of the protein films. SC3 shows a rather homogenous coverage of the mica surface, with fibrillary structures. Multiple compressions to a surface pressure of 13 mn m-1 led to a shortening of the fibrils. HFBII exhibits multilayered structures of varying thickness at higher surface pressures. Multiple compressions led to a variety of large, multilayer aggregates. Several compressions and expansions homogenized the films of both types. Both proteins showed similar dendritic structures with relevant length scales of at least several hundred nanometers at pressures of 13 mn m-1 and above, although the primary structures they assemble into are usually different in size and type, and range from fibrils to hexagonally ordered films. These dendritic structures may stem from a combination of mechanical influences, such as compressions, expansions, and the drying effect during LS transfer, which may simulate processes during physiological applications of hydrophobins, such as encapsulation or release of spores.