2BV9

HOW FAMILY 26 GLYCOSIDE HYDROLASES ORCHESTRATE CATALYSIS ON DIFFERENT POLYSACCHARIDES. STRUCTURE AND ACTIVITY OF A CLOSTRIDIUM THERMOCELLUM LICHENASE, CtLIC26A

2BV9 の概要

• エントリーDOI: 10.2210/pdb2bv9/pdb
• 関連するPDBエントリー: 1V0A 2BVD
• 分子名称: ENDOGLUCANASE H (2 entities in total)
• 機能のキーワード: hydrolase, beta-1 4 beta-1 3 glucanase, glycoside hydrolase family 26
• 由来する生物種: CLOSTRIDIUM THERMOCELLUM
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 33188.63

構造登録者

Taylor, E.J.,Goyal, A.,Guerreiro, C.I.P.D.,Prates, J.A.M.,Money, V.A.,Ferry, N.,Morland, C.,Planas, A.,Macdonald, J.A.,Stick, R.V.,Gilbert, H.J.,Fontes, C.M.G.A.,Davies, G.J. (登録日: 2005-06-23, 公開日: 2005-06-30, 最終更新日: 2024-05-08)

主引用文献

Taylor, E.J.,Goyal, A.,Guerreiro, C.I.P.D.,Prates, J.A.M.,Money, V.A.,Ferry, N.,Morland, C.,Planas, A.,Macdonald, J.A.,Stick, R.V.,Gilbert, H.J.,Fontes, C.M.G.A.,Davies, G.J.
How Family 26 Glycoside Hydrolases Orchestrate Catalysis on Different Polysaccharides: Structure and Activity of a Clostridium Thermocellum Lichenase, Ctlic26A.
J.Biol.Chem., 280:32761-, 2005

PubMed Abstract: One of the most intriguing features of the 90 glycoside hydrolase families (GHs) is the range of specificities displayed by different members of the same family, whereas the catalytic apparatus and mechanism are often invariant. Family GH26 predominantly comprises beta-1,4 mannanases; however, a bifunctional Clostridium thermocellum GH26 member (hereafter CtLic26A) displays a markedly different specificity. We show that CtLic26A is a lichenase, specific for mixed (Glcbeta1,4Glcbeta1,4Glcbeta1,3)n oligo- and polysaccharides, and displays no activity on manno-configured substrates or beta-1,4-linked homopolymers of glucose or xylose. The three-dimensional structure of the native form of CtLic26A has been solved at 1.50-A resolution, revealing a characteristic (beta/alpha)8 barrel with Glu-109 and Glu-222 acting as the catalytic acid/base and nucleophile in a double-displacement mechanism. The complex with the competitive inhibitor, Glc-beta-1,3-isofagomine (Ki 1 microm), at 1.60 A sheds light on substrate recognition in the -2 and -1 subsites and illuminates why the enzyme is specific for lichenan-based substrates. Hydrolysis of beta-mannosides by GH26 members is thought to proceed through transition states in the B2,5 (boat) conformation in which structural distinction of glucosides versus mannosides reflects not the configuration at C2 but the recognition of the pseudoaxial O3 of the B2,5 conformation. We suggest a different conformational itinerary for the GH26 enzymes active on gluco-configured substrates.

PubMed: 15987675
DOI: 10.1074/JBC.M506580200

実験手法

X-RAY DIFFRACTION (1.5 Å)