Structure-function relationships in hydrophobins: Probing the role of charged side chains

M. Lienemann, J.-A. Gandier, J. Joensuu, A. Iwanaga, Y. Takatsuji, T. Haruyama, E. Master, M. Tenkanen, M. B. Linder

Research output: Contribution to journalArticleScientificpeer-review

13 Citations (Scopus)

Abstract

Hydrophobins are small fungal proteins that are amphiphilic and have a strong tendency to assemble at interfaces. By taking advantage of this property, hydrophobins have been used for a number of applications: as affinity tags in protein purification, for protein immobilization, such as in foam stabilizers, and as dispersion agents for insoluble drug molecules. Here, we used site-directed mutagenesis to gain an understanding of the molecular basis of their properties. We especially focused on the role of charged amino acids in the structure of hydrophobins. For this purpose, fusion proteins consisting of Trichoderma reesei hydrophobin I (HFBI) and the green fluorescent protein (GFP) that contained various combinations of substitutions of charged amino acids (D30, K32, D40, D43, R45, K50) in the HFBI structure were produced. The effects of the introduced mutations on binding, oligomerization, and partitioning were characterized in an aqueous two-phase system. It was found that some substitutions caused better surface binding and reduced oligomerization, while some showed the opposite effects. However, all mutations decreased partitioning in surfactant systems, indicating that the different functions are not directly correlated and that partitioning is dependent on finely tuned properties of hydrophobins. This work shows that not all functions in self-assembly are connected in a predictable way and that a simple surfactant model for hydrophobin function is insufficient.
Original languageEnglish
Pages (from-to)5533 - 5538
JournalApplied and Environmental Microbiology
Volume79
Issue number18
DOIs
Publication statusPublished - 8 Jul 2013
MoE publication typeA1 Journal article-refereed

Fingerprint

structure-activity relationships
Surface-Active Agents
protein
partitioning
Mutation
Proteins
Trichoderma
Fungal Proteins
surfactant
Amino Acid Substitution
Site-Directed Mutagenesis
mutation
Green Fluorescent Proteins
substitution
surfactants
Immobilization
amino acid
fungal proteins
Amino Acids
Trichoderma reesei

Cite this

Lienemann, M. ; Gandier, J.-A. ; Joensuu, J. ; Iwanaga, A. ; Takatsuji, Y. ; Haruyama, T. ; Master, E. ; Tenkanen, M. ; Linder, M. B. / Structure-function relationships in hydrophobins : Probing the role of charged side chains. In: Applied and Environmental Microbiology. 2013 ; Vol. 79, No. 18. pp. 5533 - 5538.
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Lienemann, M, Gandier, J-A, Joensuu, J, Iwanaga, A, Takatsuji, Y, Haruyama, T, Master, E, Tenkanen, M & Linder, MB 2013, 'Structure-function relationships in hydrophobins: Probing the role of charged side chains', Applied and Environmental Microbiology, vol. 79, no. 18, pp. 5533 - 5538. https://doi.org/10.1128/AEM.01493-13

Structure-function relationships in hydrophobins : Probing the role of charged side chains. / Lienemann, M.; Gandier, J.-A.; Joensuu, J.; Iwanaga, A.; Takatsuji, Y.; Haruyama, T.; Master, E.; Tenkanen, M.; Linder, M. B.

In: Applied and Environmental Microbiology, Vol. 79, No. 18, 08.07.2013, p. 5533 - 5538.

Research output: Contribution to journalArticleScientificpeer-review

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T2 - Probing the role of charged side chains

AU - Lienemann, M.

AU - Gandier, J.-A.

AU - Joensuu, J.

AU - Iwanaga, A.

AU - Takatsuji, Y.

AU - Haruyama, T.

AU - Master, E.

AU - Tenkanen, M.

AU - Linder, M. B.

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Y1 - 2013/7/8

N2 - Hydrophobins are small fungal proteins that are amphiphilic and have a strong tendency to assemble at interfaces. By taking advantage of this property, hydrophobins have been used for a number of applications: as affinity tags in protein purification, for protein immobilization, such as in foam stabilizers, and as dispersion agents for insoluble drug molecules. Here, we used site-directed mutagenesis to gain an understanding of the molecular basis of their properties. We especially focused on the role of charged amino acids in the structure of hydrophobins. For this purpose, fusion proteins consisting of Trichoderma reesei hydrophobin I (HFBI) and the green fluorescent protein (GFP) that contained various combinations of substitutions of charged amino acids (D30, K32, D40, D43, R45, K50) in the HFBI structure were produced. The effects of the introduced mutations on binding, oligomerization, and partitioning were characterized in an aqueous two-phase system. It was found that some substitutions caused better surface binding and reduced oligomerization, while some showed the opposite effects. However, all mutations decreased partitioning in surfactant systems, indicating that the different functions are not directly correlated and that partitioning is dependent on finely tuned properties of hydrophobins. This work shows that not all functions in self-assembly are connected in a predictable way and that a simple surfactant model for hydrophobin function is insufficient.

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U2 - 10.1128/AEM.01493-13

DO - 10.1128/AEM.01493-13

M3 - Article

VL - 79

SP - 5533

EP - 5538

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

IS - 18

ER -