6IXD

X-ray crystal structure of bPI-11 hiv-1 protease complex

6IXD の概要

• エントリーDOI: 10.2210/pdb6ixd/pdb
• 分子名称: Protease, GLYCEROL, SULFATE ION, ... (6 entities in total)
• 機能のキーワード: hiv-1 protease, hydrolase, hydrolase-hydrolase inhibitor complex, hydrolase/hydrolase inhibitor
• 由来する生物種: Human immunodeficiency virus 1
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 22858.31

構造登録者

Adachi, M.,Hidaka, K. (登録日: 2018-12-10, 公開日: 2019-05-22, 最終更新日: 2023-11-22)

主引用文献

Hidaka, K.,Adachi, M.,Tsuda, Y.
Acquired Removability of Aspartic Protease Inhibitors by Direct Biotinylation.
Bioconjug.Chem., 30:1979-1985, 2019

PubMed Abstract: Protease inhibitors are used as both research tools and therapeutics. Many of these inhibitors consist of substrate amino acid sequence-derived structure with a transition state mimic to interact with the active site of the protease, suppressing enzymatic activity. However, once they bind, macrodilution or protein denaturation is required to remove them, limiting their usage. In this study, we describe a removable protease inhibitor, which is a directly biotinylated analogue to control the activities of HIV-1 protease and human cathepsin D. In the substrate cleavage assay, we observed that the nanomolar inhibitory activities were lost upon the addition of streptavidin, while the enzymatic activities sufficiently recovered. HIV-1 protease mixed with the removable inhibitor, avoiding autolysis, was still active to be detected by adding streptavidin after one year at room temperature. We also observed that the inhibitor was an effective eluent for the simple detection of the activity of proteases purified from human serum and cells. These results demonstrate that direct biotinylation of protease inhibitors could be a novel method for controlling the enzymatic activity from OFF to ON. We proposed the phenomenon that binding equilibrium of inhibitor was shifted from protease to streptavidin with higher affinity, named "inhibitor stripping action by affinity competition", or ISAAC. We anticipate that ISAAC could be applicable for preservatives of proteases and activity-based diagnosis of protease related diseases. Furthermore, removable inhibitor to be designed for targeted proteases changing the inhibitor structure may elucidate enzymatic activity in intrinsic form with natural modifications from various biological samples.

PubMed: 30990716
DOI: 10.1021/acs.bioconjchem.9b00195

実験手法

X-RAY DIFFRACTION (1 Å)