1W3L

ENDOGLUCANASE CEL5A FROM BACILLUS AGARADHAERENS IN COMPLEX WITH CELLOTRI DERIVED-TETRAHYDROOXAZINE

1W3L の概要

• エントリーDOI: 10.2210/pdb1w3l/pdb
• 関連するPDBエントリー: 1A3H 1H11 1H2J 1H5V 1HF6 1OCQ 1W3J 1W3K 2A3H 3A3H 5A3H 6A3H 7A3H
• 関連するBIRD辞書のPRD_ID: PRD_900005
• 分子名称: ENDOGLUCANASE 5A, beta-D-glucopyranose-(1-4)-beta-D-glucopyranose, GLYCEROL, ... (6 entities in total)
• 機能のキーワード: hydrolase, glycoside hydrolase, cellulose degradation, endoglucanase, family 5, tetrahydrooxazine
• 由来する生物種: BACILLUS AGARADHAERENS
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 34769.72

構造登録者

Gloster, T.M.,Macdonald, J.M.,Tarling, C.A.,Stick, R.V.,Withers, S.W.,Davies, G.J. (登録日: 2004-07-16, 公開日: 2004-09-08, 最終更新日: 2025-10-01)

主引用文献

Gloster, T.M.,Macdonald, J.M.,Tarling, C.A.,Stick, R.V.,Withers, S.W.,Davies, G.J.
Structural, Thermodynamic, and Kinetic Analyses of Tetrahydrooxazine-Derived Inhibitors Bound to {Beta}-Glucosidases
J.Biol.Chem., 279:49236-, 2004

PubMed Abstract: The understanding of transition state mimicry in glycoside hydrolysis is increasingly important both in the quest for novel specific therapeutic agents and for the deduction of enzyme function and mechanism. To aid comprehension, inhibitors can be characterized through kinetic, thermodynamic, and structural dissection to build an "inhibition profile." Here we dissect the binding of a tetrahydrooxazine inhibitor and its derivatives, which display Ki values around 500 nm. X-ray structures with both a beta-glucosidase, at 2 A resolution, and an endoglucanase at atomic (approximately 1 A) resolution reveal similar interactions between the tetrahydrooxazine inhibitor and both enzymes. Kinetic analyses reveal the pH dependence of kcat/Km and 1/Ki with both enzyme systems, and isothermal titration calorimetry unveils the enthalpic and entropic contributions to beta-glucosidase inhibition. The pH dependence of enzyme activity mirrored that of 1/Ki in both enzymes, unlike the cases of isofagomine and 1-deoxynojirimycin that have been characterized previously. Calorimetric dissection reveals a large favorable enthalpy that is partially offset by an unfavorable entropy upon binding. In terms of the similar profile for the pH dependence of 1/Ki and the pH dependence of kcat/Km, the significant enthalpy of binding when compared with other glycosidase inhibitors, and the tight binding at the optimal pH of the enzymes tested, tetrahydrooxazine and its derivatives are a significantly better class of glycosidase inhibitor than previously assumed.

PubMed: 15356002
DOI: 10.1074/JBC.M407195200

実験手法

X-RAY DIFFRACTION (1.04 Å)