2WJ1

3D-crystal structure of humanized-rat fatty acid amide hydrolase (FAAH) conjugated with 7-phenyl-1-(4-(pyridin-2-yl)oxazol-2-yl)heptan- 1-one, an alpha-ketooxazole

2WJ1 の概要

• エントリーDOI: 10.2210/pdb2wj1/pdb
• 関連するPDBエントリー: 1MT5 2VYA 2WAP
• 分子名称: FATTY-ACID AMIDE HYDROLASE 1, 7-phenyl-1-(4-pyridin-2-yl-1,3-oxazol-2-yl)heptane-1,1-diol, CHLORIDE ION, ... (4 entities in total)
• 機能のキーワード: monotopic membrane protein, fatty acid amide hydrolase, golgi apparatus, endoplasmic reticulum, membrane, hydrolase, transmembrane, phosphoprotein, covalent reversible inhibitor
• 由来する生物種: RATTUS NORVEGICUS (RAT)
• 細胞内の位置: Endoplasmic reticulum membrane; Single-pass membrane protein: P97612
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 126963.63

構造登録者

Mileni, M.,Garfunkle, J.,DeMartino, J.K.,Cravatt, B.F.,Boger, D.L.,Stevens, R.C. (登録日: 2009-05-19, 公開日: 2009-09-15, 最終更新日: 2024-10-23)

主引用文献

Mileni, M.,Garfunkle, J.,Demartino, J.K.,Cravatt, B.F.,Boger, D.L.,Stevens, R.C.
Binding and Inactivation Mechanism of a Humanized Fatty Acid Amide Hydrolase by Alpha-Ketoheterocycle Inhibitors Revealed from Cocrystal Structures.
J.Am.Chem.Soc., 131:10497-, 2009

PubMed Abstract: The cocrystal X-ray structures of two isomeric alpha-ketooxazole inhibitors (1 (OL-135) and 2) bound to fatty acid amide hydrolase (FAAH), a key enzymatic regulator of endocannabinoid signaling, are disclosed. The active site catalytic Ser241 is covalently bound to the inhibitors' electrophilic carbonyl groups, providing the first structures of FAAH bound to an inhibitor as a deprotonated hemiketal mimicking the enzymatic tetrahedral intermediate. The work also offers a detailed view of the oxyanion hole and an exceptional "in-action" depiction of the unusual Ser-Ser-Lys catalytic triad. These structures capture the first picture of inhibitors that span the active site into the cytosolic port providing new insights that help to explain FAAH's interaction with substrate leaving groups and their role in modulating inhibitor potency and selectivity. The role for the activating central heterocycle is clearly defined and distinguished from that observed in prior applications with serine proteases, reconciling the large electronic effect of attached substituents found unique to this class of inhibitors with FAAH. Additional striking active site flexibility is seen upon binding of the inhibitors, providing insights into the existence of a now well-defined membrane access channel with the disappearance of a spatially independent portion of the acyl chain-binding pocket. Finally, comparison of the structures of OL-135 (1) and its isomer 2 indicates that they bind identically to FAAH, albeit with reversed orientations of the central activating heterocycle, revealing that the terminal 2-pyridyl substituent and the acyl chain phenyl group provide key anchoring interactions and confirming the distinguishing role of the activating oxazole.

PubMed: 19722626
DOI: 10.1021/JA902694N

実験手法

X-RAY DIFFRACTION (1.84 Å)