3B00

Crystal structure of the laminarinase catalytic domain from Thermotoga maritima MSB8 in complex with cetyltrimethylammonium bromide

Summary for 3B00

• Entry DOI: 10.2210/pdb3b00/pdb
• Related: 3AZX 3AZY 3AZZ 3B01
• Descriptor: Laminarinase, CETYL-TRIMETHYL-AMMONIUM, CALCIUM ION, ... (4 entities in total)
• Functional Keywords: beta-jelly roll fold, glycosyl hydrolase family 16, laminarinase, endo-1, 3-beta-glucanase, hydrolase
• Biological source: Thermotoga maritima
• Total number of polymer chains: 4
• Total formula weight: 125959.69

Authors

Jeng, W.Y.,Wang, N.C.,Wang, A.H.J. (deposition date: 2011-06-03, release date: 2011-11-23, Last modification date: 2023-11-01)

Primary citation

Jeng, W.Y.,Wang, N.C.,Lin, C.T.,Shyur, L.F.,Wang, A.H.
Crystal structures of the laminarinase catalytic domain from Thermotoga maritima MSB8 in complex with inhibitors: essential residues for beta-1,3 and beta-1,4 glucan selection.
J.Biol.Chem., 286:45030-45040, 2011

PubMed 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.

PubMed: 22065588
DOI: 10.1074/jbc.M111.271213

Experimental method

X-RAY DIFFRACTION (1.74 Å)