The crystal structure of the catalytic core domain of endoglucanase I from Trichoderma reesei at 3.6 Å resolution, and a comparison with related enzymes

Gerard Kleywegt, Jinyu Zou, Christina Divne, G. Davies, Irmgard Sinning, Jerry Ståhlberg, Tapani Reinikainen, Malee Srisodsuk, Tuula Teeri, T. Alwyn Jones (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

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Abstract

Cellulose is the most abundant polymer in the biosphere. Although generally resistant to degradation, it may be hydrolysed by cellulolytic organisms that have evolved a variety of structurally distinct enzymes, cellobiohydrolases and endoglucanases, for this purpose. Endoglucanase I (EG I) is the major endoglucanase produced by the cellulolytic fungus Trichoderma reesei, accounting for 5 to 10% of the total amount of cellulases produced by this organism. Together with EG I from Humicola insolens and T. reesei cellobiohydrolase I (CBH I), the enzyme is classified into family 7 of the glycosyl hydrolases, and it catalyses hydrolysis with a net retention of the anomeric configuration.

The structure of the catalytic core domain (residues 1 to 371) of EG I from T. reesei has been determined at 3.6 Å resolution by the molecular replacement method using the structures of T. reesei CBH I and H. insolens EG I as search models. By employing the 2-fold non-crystallographic symmetry (NCS), the structure was refined successfully, despite the limited resolution. The final model has an R-factor of 0.201 (Rfree 0.258).

The structure of EG I reveals an extended, open substrate-binding cleft, rather than a tunnel as found in the homologous cellobiohydrolase CBH I. This confirms the earlier proposal that the tunnel-forming loops in CBH I have been deleted in EG I, which has resulted in an open active site in EG I, enabling it to function as an endoglucanase. Comparison of the structure of EG I with several related enzymes reveals structural similarities, and differences that relate to their biological function in degrading particular substrates. A possible structural explanation of the drastically different pH profiles of T. reesei and H. insolens EG I is proposed.

Original languageEnglish
Pages (from-to)383 - 397
Number of pages15
JournalJournal of Molecular Biology
Volume272
Issue number3
DOIs
Publication statusPublished - 1997
MoE publication typeA1 Journal article-refereed

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Cellulases
Trichoderma
Catalytic Domain
Cellulose 1,4-beta-Cellobiosidase
Enzymes
Cellulase
R388
Hydrolases
Cellulose
Polymers
Hydrolysis
Fungi

Cite this

Kleywegt, Gerard ; Zou, Jinyu ; Divne, Christina ; Davies, G. ; Sinning, Irmgard ; Ståhlberg, Jerry ; Reinikainen, Tapani ; Srisodsuk, Malee ; Teeri, Tuula ; Jones, T. Alwyn. / The crystal structure of the catalytic core domain of endoglucanase I from Trichoderma reesei at 3.6 Å resolution, and a comparison with related enzymes. In: Journal of Molecular Biology. 1997 ; Vol. 272, No. 3. pp. 383 - 397.
@article{9a6bc803d353433f97eb90e04c65011e,
title = "The crystal structure of the catalytic core domain of endoglucanase I from Trichoderma reesei at 3.6 {\AA} resolution, and a comparison with related enzymes",
abstract = "Cellulose is the most abundant polymer in the biosphere. Although generally resistant to degradation, it may be hydrolysed by cellulolytic organisms that have evolved a variety of structurally distinct enzymes, cellobiohydrolases and endoglucanases, for this purpose. Endoglucanase I (EG I) is the major endoglucanase produced by the cellulolytic fungus Trichoderma reesei, accounting for 5 to 10{\%} of the total amount of cellulases produced by this organism. Together with EG I from Humicola insolens and T. reesei cellobiohydrolase I (CBH I), the enzyme is classified into family 7 of the glycosyl hydrolases, and it catalyses hydrolysis with a net retention of the anomeric configuration.The structure of the catalytic core domain (residues 1 to 371) of EG I from T. reesei has been determined at 3.6 {\AA} resolution by the molecular replacement method using the structures of T. reesei CBH I and H. insolens EG I as search models. By employing the 2-fold non-crystallographic symmetry (NCS), the structure was refined successfully, despite the limited resolution. The final model has an R-factor of 0.201 (Rfree 0.258).The structure of EG I reveals an extended, open substrate-binding cleft, rather than a tunnel as found in the homologous cellobiohydrolase CBH I. This confirms the earlier proposal that the tunnel-forming loops in CBH I have been deleted in EG I, which has resulted in an open active site in EG I, enabling it to function as an endoglucanase. Comparison of the structure of EG I with several related enzymes reveals structural similarities, and differences that relate to their biological function in degrading particular substrates. A possible structural explanation of the drastically different pH profiles of T. reesei and H. insolens EG I is proposed.",
author = "Gerard Kleywegt and Jinyu Zou and Christina Divne and G. Davies and Irmgard Sinning and Jerry St{\aa}hlberg and Tapani Reinikainen and Malee Srisodsuk and Tuula Teeri and Jones, {T. Alwyn}",
year = "1997",
doi = "10.1006/jmbi.1997.1243",
language = "English",
volume = "272",
pages = "383 -- 397",
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Kleywegt, G, Zou, J, Divne, C, Davies, G, Sinning, I, Ståhlberg, J, Reinikainen, T, Srisodsuk, M, Teeri, T & Jones, TA 1997, 'The crystal structure of the catalytic core domain of endoglucanase I from Trichoderma reesei at 3.6 Å resolution, and a comparison with related enzymes', Journal of Molecular Biology, vol. 272, no. 3, pp. 383 - 397. https://doi.org/10.1006/jmbi.1997.1243

The crystal structure of the catalytic core domain of endoglucanase I from Trichoderma reesei at 3.6 Å resolution, and a comparison with related enzymes. / Kleywegt, Gerard; Zou, Jinyu; Divne, Christina; Davies, G.; Sinning, Irmgard; Ståhlberg, Jerry; Reinikainen, Tapani; Srisodsuk, Malee; Teeri, Tuula; Jones, T. Alwyn (Corresponding Author).

In: Journal of Molecular Biology, Vol. 272, No. 3, 1997, p. 383 - 397.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - The crystal structure of the catalytic core domain of endoglucanase I from Trichoderma reesei at 3.6 Å resolution, and a comparison with related enzymes

AU - Kleywegt, Gerard

AU - Zou, Jinyu

AU - Divne, Christina

AU - Davies, G.

AU - Sinning, Irmgard

AU - Ståhlberg, Jerry

AU - Reinikainen, Tapani

AU - Srisodsuk, Malee

AU - Teeri, Tuula

AU - Jones, T. Alwyn

PY - 1997

Y1 - 1997

N2 - Cellulose is the most abundant polymer in the biosphere. Although generally resistant to degradation, it may be hydrolysed by cellulolytic organisms that have evolved a variety of structurally distinct enzymes, cellobiohydrolases and endoglucanases, for this purpose. Endoglucanase I (EG I) is the major endoglucanase produced by the cellulolytic fungus Trichoderma reesei, accounting for 5 to 10% of the total amount of cellulases produced by this organism. Together with EG I from Humicola insolens and T. reesei cellobiohydrolase I (CBH I), the enzyme is classified into family 7 of the glycosyl hydrolases, and it catalyses hydrolysis with a net retention of the anomeric configuration.The structure of the catalytic core domain (residues 1 to 371) of EG I from T. reesei has been determined at 3.6 Å resolution by the molecular replacement method using the structures of T. reesei CBH I and H. insolens EG I as search models. By employing the 2-fold non-crystallographic symmetry (NCS), the structure was refined successfully, despite the limited resolution. The final model has an R-factor of 0.201 (Rfree 0.258).The structure of EG I reveals an extended, open substrate-binding cleft, rather than a tunnel as found in the homologous cellobiohydrolase CBH I. This confirms the earlier proposal that the tunnel-forming loops in CBH I have been deleted in EG I, which has resulted in an open active site in EG I, enabling it to function as an endoglucanase. Comparison of the structure of EG I with several related enzymes reveals structural similarities, and differences that relate to their biological function in degrading particular substrates. A possible structural explanation of the drastically different pH profiles of T. reesei and H. insolens EG I is proposed.

AB - Cellulose is the most abundant polymer in the biosphere. Although generally resistant to degradation, it may be hydrolysed by cellulolytic organisms that have evolved a variety of structurally distinct enzymes, cellobiohydrolases and endoglucanases, for this purpose. Endoglucanase I (EG I) is the major endoglucanase produced by the cellulolytic fungus Trichoderma reesei, accounting for 5 to 10% of the total amount of cellulases produced by this organism. Together with EG I from Humicola insolens and T. reesei cellobiohydrolase I (CBH I), the enzyme is classified into family 7 of the glycosyl hydrolases, and it catalyses hydrolysis with a net retention of the anomeric configuration.The structure of the catalytic core domain (residues 1 to 371) of EG I from T. reesei has been determined at 3.6 Å resolution by the molecular replacement method using the structures of T. reesei CBH I and H. insolens EG I as search models. By employing the 2-fold non-crystallographic symmetry (NCS), the structure was refined successfully, despite the limited resolution. The final model has an R-factor of 0.201 (Rfree 0.258).The structure of EG I reveals an extended, open substrate-binding cleft, rather than a tunnel as found in the homologous cellobiohydrolase CBH I. This confirms the earlier proposal that the tunnel-forming loops in CBH I have been deleted in EG I, which has resulted in an open active site in EG I, enabling it to function as an endoglucanase. Comparison of the structure of EG I with several related enzymes reveals structural similarities, and differences that relate to their biological function in degrading particular substrates. A possible structural explanation of the drastically different pH profiles of T. reesei and H. insolens EG I is proposed.

U2 - 10.1006/jmbi.1997.1243

DO - 10.1006/jmbi.1997.1243

M3 - Article

VL - 272

SP - 383

EP - 397

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

IS - 3

ER -