Behavior of Trichoderma reesei hydrophobins in solution: interactions, dynamics, and multimer formation.
Filamentous fungi utilize small amphiphilic proteins called hydrophobins in their adaptation to the environment. The hydrophobins are used to form coatings on various fungal structures, lower the surface tension of water, and to mediate surface attachment. Hydrophobins function through self-assembly at interfaces, for example, at the air-water interface, and at fungal cellular structures. Despite their high tendency to self assemble at interfaces, hydrophobins can be very soluble in water. To understand the mechanism of hydrophobin self-assembly, in this work, we have studied the behavior of two Trichoderma reesei hydrophobins, HFBI and HFBII in aqueous solution. The main methods used were Förster resonance energy transfer (FRET) and size exclusion chromatography. A genetically engineered HFBI variant, NCys-HFBI, was utilized for the site-specific labeling of dyes for the FRET experiments. We observed the multimerization of HFBI in a concentration-dependent manner. A change from monomers to tetramers was seen when the hydrophobin concentration was increased. Interaction studies between HFBI and HFBII suggested that at low concentrations homodimers are preferred, and at higher concentrations, the heterotetramers of HFBI and HFBII are formed. In conclusion, the results support the model where hydrophobins in aqueous solutions form multimers by hydrophobic interactions. In contrast to micelles formed by detergents, the hydrophobin multimers are defined in size and involve specific protein-protein interactions.
Szilvay
Géza R
GR
VTT Biotechnology, Tietotie 2, P.O. Box 1000, FIN-02044 VTT, Finland. geza.szilvay@vtt.fi
Nakari-Setälä
Tiina
T
Linder
Markus B
MB
D018360:Crystallography, X-Ray; D003545:Cysteine; D031541:Fluorescence Resonance Energy Transfer; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics; D011487:Protein Conformation; D012996:Solutions / Q000737:chemistry; D014242:Trichoderma / Q000378:metabolism*
D005656:Fungal Proteins; D012996:Solutions; D003545:Cysteine