4INH

Structural Basis of Substrate Specificity and Protease Inhibition in Norwalk Virus

4INH の概要

• エントリーDOI: 10.2210/pdb4inh/pdb
• 関連するPDBエントリー: 4IMQ 4IMZ 4IN1 4IN2
• 関連するBIRD辞書のPRD_ID: PRD_001054
• 分子名称: Genome polyprotein, peptide inhibitor, syc59, DIMETHYL SULFOXIDE, ... (4 entities in total)
• 機能のキーワード: protease, hydrolase-hydrolase inhibitor complex, hydrolase/hydrolase inhibitor
• 由来する生物種: Norovirus Hu/1968/US (Hu/NV/NV/1968/US); 詳細
• 細胞内の位置: Protein p48: Host membrane; Single-pass membrane protein (Potential). NTPase: Host membrane; Single-pass membrane protein (Potential). Protein p22: Host membrane; Single-pass membrane protein (Potential): Q83883
• タンパク質・核酸の鎖数: 16
• 化学式量合計: 161412.70

構造登録者

Prasad, B.V.V.,Muhaxhiri, Z.,Deng, L.,Shanker, S.,Sankaran, B.,Estes, M.K.,Palzkill, T.,Song, Y. (登録日: 2013-01-04, 公開日: 2013-02-20, 最終更新日: 2025-03-26)

主引用文献

Muhaxhiri, Z.,Deng, L.,Shanker, S.,Sankaran, B.,Estes, M.K.,Palzkill, T.,Song, Y.,Prasad, B.V.
Structural basis of substrate specificity and protease inhibition in norwalk virus.
J.Virol., 87:4281-4292, 2013

PubMed Abstract: Norwalk virus (NV), the prototype human calicivirus, is the leading cause of nonbacterial acute gastroenteritis. The NV protease cleaves the polyprotein encoded by open reading frame 1 of the viral genome at five nonhomologous sites, releasing six nonstructural proteins that are essential for viral replication. The structural details of how NV protease recognizes multiple substrates are unclear. In our X-ray structure of an NV protease construct, we observed that the C-terminal tail, representing the native substrate positions P5 to P1, is inserted into the active site cleft of the neighboring protease molecule, providing atomic details of how NV protease recognizes a substrate. The crystallographic structure of NV protease with the C-terminal tail redesigned to mimic P4 to P1 of another substrate site provided further structural details on how the active site accommodates sequence variations in the substrates. Based on these structural analyses, substrate-based aldehyde inhibitors were synthesized and screened for inhibition potency. Crystallographic structures of the protease in complex with each of the three most potent inhibitors were determined. These structures showed concerted conformational changes in the S4 and S2 pockets of the protease to accommodate variations in the P4 and P2 residues of the substrate/inhibitor, which could be a mechanism for how the NV protease recognizes multiple sites in the polyprotein with differential affinities during virus replication. These structures further indicate that the mechanism of inhibition by these inhibitors involves covalent bond formation with the side chain of the conserved cysteine in the active site by nucleophilic addition, and such substrate-based aldehydes could be effective protease inhibitors.

PubMed: 23365454
DOI: 10.1128/JVI.02869-12

実験手法

X-RAY DIFFRACTION (1.7 Å)