The binding specificity and affinity determinants of family 1 and family 3 cellulose binding modules

Janne Lehtiö, Junji Sugiyama, Malin Gustavsson, Linda Fransson, Markus Linder, Tuula Teeri (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

247 Citations (Scopus)

Abstract

Cellulose binding modules (CBMs) potentiate the action of cellulolytic enzymes on insoluble substrates. Numerous studies have established that three aromatic residues on a CBM surface are needed for binding onto cellulose crystals and that tryptophans contribute to higher binding affinity than tyrosines. However, studies addressing the nature of CBM–cellulose interactions have so far failed to establish the binding site on cellulose crystals targeted by CBMs. In this study, the binding sites of CBMs on Valonia cellulose crystals have been visualized by transmission electron microscopy. Fusion of the CBMs with a modified staphylococcal protein A (ZZ-domain) allowed direct immuno-gold labeling at close proximity of the actual CBM binding site. The transmission electron microscopy images provide unequivocal evidence that the fungal family 1 CBMs as well as the family 3 CBM from Clostridium thermocellum CipA have defined binding sites on two opposite corners of Valonia cellulose crystals. In most samples these corners are worn to display significant area of the hydrophobic (110) plane, which thus constitutes the binding site for these CBMs.
Original languageEnglish
Pages (from-to)484-489
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume100
Issue number2
DOIs
Publication statusPublished - 2003
MoE publication typeA1 Journal article-refereed

Fingerprint

Cellulose
Binding Sites
Crystals
Transmission electron microscopy
Clostridium
Staphylococcal Protein A
Tryptophan
Gold
Labeling
Tyrosine
Fusion reactions

Keywords

  • cellulose
  • cellulose binding
  • cellulose-binding domain

Cite this

Lehtiö, Janne ; Sugiyama, Junji ; Gustavsson, Malin ; Fransson, Linda ; Linder, Markus ; Teeri, Tuula. / The binding specificity and affinity determinants of family 1 and family 3 cellulose binding modules. In: Proceedings of the National Academy of Sciences of the United States of America. 2003 ; Vol. 100, No. 2. pp. 484-489.
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abstract = "Cellulose binding modules (CBMs) potentiate the action of cellulolytic enzymes on insoluble substrates. Numerous studies have established that three aromatic residues on a CBM surface are needed for binding onto cellulose crystals and that tryptophans contribute to higher binding affinity than tyrosines. However, studies addressing the nature of CBM–cellulose interactions have so far failed to establish the binding site on cellulose crystals targeted by CBMs. In this study, the binding sites of CBMs on Valonia cellulose crystals have been visualized by transmission electron microscopy. Fusion of the CBMs with a modified staphylococcal protein A (ZZ-domain) allowed direct immuno-gold labeling at close proximity of the actual CBM binding site. The transmission electron microscopy images provide unequivocal evidence that the fungal family 1 CBMs as well as the family 3 CBM from Clostridium thermocellum CipA have defined binding sites on two opposite corners of Valonia cellulose crystals. In most samples these corners are worn to display significant area of the hydrophobic (110) plane, which thus constitutes the binding site for these CBMs.",
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The binding specificity and affinity determinants of family 1 and family 3 cellulose binding modules. / Lehtiö, Janne; Sugiyama, Junji; Gustavsson, Malin; Fransson, Linda; Linder, Markus; Teeri, Tuula (Corresponding Author).

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 100, No. 2, 2003, p. 484-489.

Research output: Contribution to journalArticleScientificpeer-review

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T1 - The binding specificity and affinity determinants of family 1 and family 3 cellulose binding modules

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AU - Sugiyama, Junji

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AU - Teeri, Tuula

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N2 - Cellulose binding modules (CBMs) potentiate the action of cellulolytic enzymes on insoluble substrates. Numerous studies have established that three aromatic residues on a CBM surface are needed for binding onto cellulose crystals and that tryptophans contribute to higher binding affinity than tyrosines. However, studies addressing the nature of CBM–cellulose interactions have so far failed to establish the binding site on cellulose crystals targeted by CBMs. In this study, the binding sites of CBMs on Valonia cellulose crystals have been visualized by transmission electron microscopy. Fusion of the CBMs with a modified staphylococcal protein A (ZZ-domain) allowed direct immuno-gold labeling at close proximity of the actual CBM binding site. The transmission electron microscopy images provide unequivocal evidence that the fungal family 1 CBMs as well as the family 3 CBM from Clostridium thermocellum CipA have defined binding sites on two opposite corners of Valonia cellulose crystals. In most samples these corners are worn to display significant area of the hydrophobic (110) plane, which thus constitutes the binding site for these CBMs.

AB - Cellulose binding modules (CBMs) potentiate the action of cellulolytic enzymes on insoluble substrates. Numerous studies have established that three aromatic residues on a CBM surface are needed for binding onto cellulose crystals and that tryptophans contribute to higher binding affinity than tyrosines. However, studies addressing the nature of CBM–cellulose interactions have so far failed to establish the binding site on cellulose crystals targeted by CBMs. In this study, the binding sites of CBMs on Valonia cellulose crystals have been visualized by transmission electron microscopy. Fusion of the CBMs with a modified staphylococcal protein A (ZZ-domain) allowed direct immuno-gold labeling at close proximity of the actual CBM binding site. The transmission electron microscopy images provide unequivocal evidence that the fungal family 1 CBMs as well as the family 3 CBM from Clostridium thermocellum CipA have defined binding sites on two opposite corners of Valonia cellulose crystals. In most samples these corners are worn to display significant area of the hydrophobic (110) plane, which thus constitutes the binding site for these CBMs.

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