4PSR

Crystal Structure of alpha-L-fucosidase from Fusarium graminearum in the open form in complex with L-fucose

4PSR の概要

• エントリーDOI: 10.2210/pdb4psr/pdb
• 関連するPDBエントリー: 3EUB 3NVW 3NVY 3NVZ 4ni3 4psp
• 分子名称: Alpha-fucosidase GH29, alpha-D-mannopyranose-(1-2)-alpha-D-mannopyranose-(1-2)-alpha-D-mannopyranose-(1-3)-[alpha-D-mannopyranose-(1-6)]beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, alpha-D-mannopyranose-(1-2)-alpha-D-mannopyranose-(1-2)-alpha-D-mannopyranose-(1-3)-beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, ... (9 entities in total)
• 機能のキーワード: fucosidase, gh29, glycoside hydrolase, tim barrel, n-glycosylation, hydrolase
• 由来する生物種: Fusarium graminearum (Wheat head blight fungus)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 135889.77

構造登録者

Cao, H.,Walton, J.,Brumm, P.,Phillips Jr., G.N. (登録日: 2014-03-07, 公開日: 2014-03-19, 最終更新日: 2024-10-30)

主引用文献

Cao, H.,Walton, J.D.,Brumm, P.,Phillips, G.N.
Structure and Substrate Specificity of a Eukaryotic Fucosidase from Fusarium graminearum.
J.Biol.Chem., 289:25624-25638, 2014

PubMed Abstract: The secreted glycoside hydrolase family 29 (GH29) α-L-fucosidase from plant pathogenic fungus Fusarium graminearum (FgFCO1) actively releases fucose from the xyloglucan fragment. We solved crystal structures of two active-site conformations, i.e. open and closed, of apoFgFCO1 and an open complex with product fucose at atomic resolution. The closed conformation supports catalysis by orienting the conserved general acid/base Glu-288 nearest the predicted glycosidic position, whereas the open conformation possibly represents an unreactive state with Glu-288 positioned away from the catalytic center. A flexible loop near the substrate binding site containing a non-conserved GGSFT sequence is ordered in the closed but not the open form. We also identified a novel C-terminal βγ-crystallin domain in FgFCO1 devoid of calcium binding motif whose homologous sequences are present in various glycoside hydrolase families. N-Glycosylated FgFCO1 adopts a monomeric state as verified by solution small angle x-ray scattering in contrast to reported multimeric fucosidases. Steady-state kinetics shows that FgFCO1 prefers α1,2 over α1,3/4 linkages and displays minimal activity with p-nitrophenyl fucoside with an acidic pH optimum of 4.6. Despite a retaining GH29 family fold, the overall specificity of FgFCO1 most closely resembles inverting GH95 α-fucosidase, which displays the highest specificity with two natural substrates harboring the Fucα1-2Gal glycosidic linkage, a xyloglucan-derived nonasaccharide, and 2'-fucosyllactose. Furthermore, FgFCO1 hydrolyzes H-disaccharide (lacking a +2 subsite sugar) at a rate 10(3)-fold slower than 2'-fucosyllactose. We demonstrated the structurally dynamic active site of FgFCO1 with flexible general acid/base Glu, a common feature shared by several bacterial GH29 fucosidases to various extents.

PubMed: 25086049
DOI: 10.1074/jbc.M114.583286

実験手法

X-RAY DIFFRACTION (1.38 Å)