Crystal structures of the laminarinase catalytic domain from Thermotoga maritima MSB8 in complex with inhibitors: Essential residues for β-1,3- and β-1,4-glucan selection

Wen-Yih Jeng, Nai Chen Wang, Cheng Tse Lin, Lie Fen Shyur, Andrew H.J. Wang

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Laminarinases hydrolyzing the β-1,3-linkage of glucans play essential roles in microbial saccharide degradation. Here we report the crystal structures at 1.65-1.82 Å resolution of the catalytic domain of laminarinase from the thermophile Thermotoga maritima with various space groups in the ligand-free form or in the presence of inhibitors gluconolactone and cetyltrimethylammonium. Ligands were bound at the cleft of the active site near an enclosure formed by Trp-232 and a flexible GASIG loop. A closed configuration at the active site cleft was observed in some molecules. The loop flexibility in the enzyme may contribute to the regulation of endo- or exo-activity of the enzyme and a preference to release laminaritrioses in long chain carbohydrate hydrolysis. Glu-137 and Glu-132 are proposed to serve as the proton donor and nucleophile, respectively, in the retaining catalysis of hydrolyzation. Calcium ions in the crystallization media are found to accelerate crystal growth. Comparison of laminarinase and endoglucanase structures revealed the subtle difference of key residues in the active site for the selection of β-1,3-glucan and β-1,4-glucan substrates, respectively. Arg-85 may be pivotal to β-1,3-glucan substrate selection. The similarity of the structures between the laminarinase catalytic domain and its carbohydrate-binding modules may have evolutionary relevance because of the similarities in their folds.

Original languageEnglish
Pages (from-to)45030-45040
Number of pages11
JournalJournal of Biological Chemistry
Volume286
Issue number52
DOIs
Publication statusPublished - 2011 Dec 30

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Thermotoga maritima
Cellulases
Catalytic Domain
Crystal structure
Glucans
Crystallization
Carbohydrates
Ligands
Nucleophiles
Cellulase
Substrates
Enzymes
Enclosures
Catalysis
Protons
Hydrolysis
Ions
Calcium
Degradation
Molecules

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

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title = "Crystal structures of the laminarinase catalytic domain from Thermotoga maritima MSB8 in complex with inhibitors: Essential residues for β-1,3- and β-1,4-glucan selection",
abstract = "Laminarinases hydrolyzing the β-1,3-linkage of glucans play essential roles in microbial saccharide degradation. Here we report the crystal structures at 1.65-1.82 {\AA} resolution of the catalytic domain of laminarinase from the thermophile Thermotoga maritima with various space groups in the ligand-free form or in the presence of inhibitors gluconolactone and cetyltrimethylammonium. Ligands were bound at the cleft of the active site near an enclosure formed by Trp-232 and a flexible GASIG loop. A closed configuration at the active site cleft was observed in some molecules. The loop flexibility in the enzyme may contribute to the regulation of endo- or exo-activity of the enzyme and a preference to release laminaritrioses in long chain carbohydrate hydrolysis. Glu-137 and Glu-132 are proposed to serve as the proton donor and nucleophile, respectively, in the retaining catalysis of hydrolyzation. Calcium ions in the crystallization media are found to accelerate crystal growth. Comparison of laminarinase and endoglucanase structures revealed the subtle difference of key residues in the active site for the selection of β-1,3-glucan and β-1,4-glucan substrates, respectively. Arg-85 may be pivotal to β-1,3-glucan substrate selection. The similarity of the structures between the laminarinase catalytic domain and its carbohydrate-binding modules may have evolutionary relevance because of the similarities in their folds.",
author = "Wen-Yih Jeng and Wang, {Nai Chen} and Lin, {Cheng Tse} and Shyur, {Lie Fen} and Wang, {Andrew H.J.}",
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Crystal structures of the laminarinase catalytic domain from Thermotoga maritima MSB8 in complex with inhibitors : Essential residues for β-1,3- and β-1,4-glucan selection. / Jeng, Wen-Yih; Wang, Nai Chen; Lin, Cheng Tse; Shyur, Lie Fen; Wang, Andrew H.J.

In: Journal of Biological Chemistry, Vol. 286, No. 52, 30.12.2011, p. 45030-45040.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Crystal structures of the laminarinase catalytic domain from Thermotoga maritima MSB8 in complex with inhibitors

T2 - Essential residues for β-1,3- and β-1,4-glucan selection

AU - Jeng, Wen-Yih

AU - Wang, Nai Chen

AU - Lin, Cheng Tse

AU - Shyur, Lie Fen

AU - Wang, Andrew H.J.

PY - 2011/12/30

Y1 - 2011/12/30

N2 - Laminarinases hydrolyzing the β-1,3-linkage of glucans play essential roles in microbial saccharide degradation. Here we report the crystal structures at 1.65-1.82 Å resolution of the catalytic domain of laminarinase from the thermophile Thermotoga maritima with various space groups in the ligand-free form or in the presence of inhibitors gluconolactone and cetyltrimethylammonium. Ligands were bound at the cleft of the active site near an enclosure formed by Trp-232 and a flexible GASIG loop. A closed configuration at the active site cleft was observed in some molecules. The loop flexibility in the enzyme may contribute to the regulation of endo- or exo-activity of the enzyme and a preference to release laminaritrioses in long chain carbohydrate hydrolysis. Glu-137 and Glu-132 are proposed to serve as the proton donor and nucleophile, respectively, in the retaining catalysis of hydrolyzation. Calcium ions in the crystallization media are found to accelerate crystal growth. Comparison of laminarinase and endoglucanase structures revealed the subtle difference of key residues in the active site for the selection of β-1,3-glucan and β-1,4-glucan substrates, respectively. Arg-85 may be pivotal to β-1,3-glucan substrate selection. The similarity of the structures between the laminarinase catalytic domain and its carbohydrate-binding modules may have evolutionary relevance because of the similarities in their folds.

AB - Laminarinases hydrolyzing the β-1,3-linkage of glucans play essential roles in microbial saccharide degradation. Here we report the crystal structures at 1.65-1.82 Å resolution of the catalytic domain of laminarinase from the thermophile Thermotoga maritima with various space groups in the ligand-free form or in the presence of inhibitors gluconolactone and cetyltrimethylammonium. Ligands were bound at the cleft of the active site near an enclosure formed by Trp-232 and a flexible GASIG loop. A closed configuration at the active site cleft was observed in some molecules. The loop flexibility in the enzyme may contribute to the regulation of endo- or exo-activity of the enzyme and a preference to release laminaritrioses in long chain carbohydrate hydrolysis. Glu-137 and Glu-132 are proposed to serve as the proton donor and nucleophile, respectively, in the retaining catalysis of hydrolyzation. Calcium ions in the crystallization media are found to accelerate crystal growth. Comparison of laminarinase and endoglucanase structures revealed the subtle difference of key residues in the active site for the selection of β-1,3-glucan and β-1,4-glucan substrates, respectively. Arg-85 may be pivotal to β-1,3-glucan substrate selection. The similarity of the structures between the laminarinase catalytic domain and its carbohydrate-binding modules may have evolutionary relevance because of the similarities in their folds.

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