Premium
Investigating the binding of β‐1,4‐galactan to Bacillus licheniformis β‐1,4‐galactanase by crystallography and computational modeling
Author(s) -
Le Nours Jérôme,
De Maria Leonardo,
Welner Ditte,
Jørgensen Christel T.,
Christensen Lars L. H.,
Borchert Torben V.,
Larsen Sine,
Lo Leggio Leila
Publication year - 2009
Publication title -
proteins: structure, function, and bioinformatics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.699
H-Index - 191
eISSN - 1097-0134
pISSN - 0887-3585
DOI - 10.1002/prot.22310
Subject(s) - galactan , glycoside hydrolase , bacillus licheniformis , cell wall , arabinogalactan , aglycone , chemistry , stereochemistry , pectin , xylobiose , polysaccharide , biochemistry , biology , glycoside , hydrolysis , xylan , bacteria , bacillus subtilis , genetics
Microbial β‐1,4‐galactanases are glycoside hydrolases belonging to family 53, which degrade galactan and arabinogalactan side chains in the hairy regions of pectin, a major plant cell wall component. They belong to the larger clan GH‐A of glycoside hydrolases, which cover many different poly‐ and oligosaccharidase specificities. Crystallographic complexes of Bacillus licheniformis β‐1,4‐galactanase and its inactive nucleophile mutant have been obtained with methyl‐β(1→4)‐galactotetraoside, providing, for the first time, information on substrate binding to the aglycone side of the β‐1,4‐galactanase substrate binding groove. Using the experimentally determined subsites as a starting point, a β(1→4)‐galactononaose was built into the structure and subjected to molecular dynamics simulations giving further insight into the residues involved in the binding of the polysaccharide from subsite −4 to +5. In particular, this analysis newly identified a conserved β‐turn, which contributes to subsites −2 to +3. This β‐turn is unique to family 53 β‐1,4‐galactanases among all clan GH‐A families that have been structurally characterized and thus might be a structural signature for endo‐β‐1,4‐galactanase specificity. Proteins 2009. © 2008 Wiley‐Liss, Inc.