5CJO

Crystal Structure Analysis of Elbow-Engineered-Fab-Bound Human Insulin Degrading Enzyme (IDE) in Complex with Insulin

5CJO の概要

• エントリーDOI: 10.2210/pdb5cjo/pdb
• 分子名称: Insulin-degrading enzyme, FAB Heavy chain with engineered elbow, FAB light chain, ... (6 entities in total)
• 機能のキーワード: hydrolase, fab, elbow-engineer, hydrolase-hydrolase inhibitor complex, hydrolase/hydrolase inhibitor
• 由来する生物種: Homo sapiens (Human); 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 166314.66

構造登録者

liang, w.g.,bailey, L.,tang, w.j. (登録日: 2015-07-14, 公開日: 2016-07-20, 最終更新日: 2024-11-06)

主引用文献

Bailey, L.J.,Sheehy, K.M.,Dominik, P.K.,Liang, W.G.,Rui, H.,Clark, M.,Jaskolowski, M.,Kim, Y.,Deneka, D.,Tang, W.J.,Kossiakoff, A.A.
Locking the Elbow: Improved Antibody Fab Fragments as Chaperones for Structure Determination.
J. Mol. Biol., 2017

PubMed Abstract: Antibody Fab fragments have been exploited with significant success to facilitate the structure determination of challenging macromolecules as crystallization chaperones and as molecular fiducial marks for single particle cryo-electron microscopy approaches. However, the inherent flexibility of the "elbow" regions, which link the constant and variable domains of the Fab, can introduce disorder and thus diminish their effectiveness. We have developed a phage display engineering strategy to generate synthetic Fab variants that significantly reduces elbow flexibility, while maintaining their high affinity and stability. This strategy was validated using previously recalcitrant Fab-antigen complexes where introduction of an engineered elbow region enhanced crystallization and diffraction resolution. Furthermore, incorporation of the mutations appears to be generally portable to other synthetic antibodies and may serve as a universal strategy to enhance the success rates of Fabs as structure determination chaperones.

PubMed: 29273204
DOI: 10.1016/j.jmb.2017.12.012

実験手法

X-RAY DIFFRACTION (3.287 Å)