Identification of functionally important amino acids in the cellulose-binding domain of Trichoderma reesei cellobiohydrolase I

Markus Linder, Maija-Liisa Mattinen (Corresponding Author), Maarit Kontteli, Gunnar Lindeberg, Jerry Ståhlberg, Torbjörn Drakenberg, Tapani Reinikainen, Göran Pettersson, Arto Annila

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192 Citations (Scopus)

Abstract

Cellobiohydrolase I (CBHI) of Trichoderma reesei has two functional domains, a catalytic core domain and a cellulose binding domain (CBD). The structure of the CBD reveals two distinct faces, one of which is flat and the other rough. Several other fungal cellulolytic enzymes have similar two‐domain structures, in which the CBDs show a conserved primary structure. Here we have evaluated the contributions of conserved amino acids in CBHI CBD to its binding to cellulose.
Binding isotherms were determined for a set of six synthetic analogues in which conserved amino acids were substituted. Two‐dimensional NMR spectroscopy was used to assess the structural effects of the substitutions by comparing chemical shifts, coupling constants, and NOEs of the backbone protons between the wild‐type CBD and the analogues. In general, the structural effects of the substitutions were minor, although in some cases decreased binding could clearly be ascribed to conformational perturbations.
We found that at least two tyrosine residues and a glutamine residue on the flat face were essential for tight binding of the CBD to cellulose. A change on the rough face had only a small effect on the binding and it is unlikely that this face interacts with cellulose directly.
Original languageEnglish
Pages (from-to)1056-1064
JournalProtein Science
Volume4
Issue number6
DOIs
Publication statusPublished - 1995
MoE publication typeA1 Journal article-refereed

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    Linder, M., Mattinen, M-L., Kontteli, M., Lindeberg, G., Ståhlberg, J., Drakenberg, T., Reinikainen, T., Pettersson, G., & Annila, A. (1995). Identification of functionally important amino acids in the cellulose-binding domain of Trichoderma reesei cellobiohydrolase I. Protein Science, 4(6), 1056-1064. https://doi.org/10.1002/pro.5560040604