2B2X

VLA1 RdeltaH I-domain complexed with a quadruple mutant of the AQC2 Fab

2B2X の概要

• エントリーDOI: 10.2210/pdb2b2x/pdb
• 関連するPDBエントリー: 1MHP
• 分子名称: Integrin alpha-1, Antibody AQC2 Fab, MAGNESIUM ION, ... (5 entities in total)
• 機能のキーワード: computational design, antibody-antigen complex, immune system
• 由来する生物種: Rattus norvegicus (Norway rat); 詳細
• 細胞内の位置: Membrane; Single-pass type I membrane protein: P18614
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 145643.02

構造登録者

Clark, L.A.,Boriack-Sjodin, P.A.,Eldredge, J.,Fitch, C.,Friedman, B.,Hanf, K.J.,Jarpe, M.,Liparoto, S.F.,Li, Y.,Lugovskoy, A. (登録日: 2005-09-19, 公開日: 2006-04-18, 最終更新日: 2024-11-13)

主引用文献

Clark, L.A.,Boriack-Sjodin, P.A.,Eldredge, J.,Fitch, C.,Friedman, B.,Hanf, K.J.,Jarpe, M.,Liparoto, S.F.,Li, Y.,Lugovskoy, A.,Miller, S.,Rushe, M.,Sherman, W.,Simon, K.,Van Vlijmen, H.
Affinity enhancement of an in vivo matured therapeutic antibody using structure-based computational design
Protein Sci., 15:949-960, 2006

PubMed Abstract: Improving the affinity of a high-affinity protein-protein interaction is a challenging problem that has practical applications in the development of therapeutic biomolecules. We used a combination of structure-based computational methods to optimize the binding affinity of an antibody fragment to the I-domain of the integrin VLA1. Despite the already high affinity of the antibody (Kd approximately 7 nM) and the moderate resolution (2.8 A) of the starting crystal structure, the affinity was increased by an order of magnitude primarily through a decrease in the dissociation rate. We determined the crystal structure of a high-affinity quadruple mutant complex at 2.2 A. The structure shows that the design makes the predicted contacts. Structural evidence and mutagenesis experiments that probe a hydrogen bond network illustrate the importance of satisfying hydrogen bonding requirements while seeking higher-affinity mutations. The large and diverse set of interface mutations allowed refinement of the mutant binding affinity prediction protocol and improvement of the single-mutant success rate. Our results indicate that structure-based computational design can be successfully applied to further improve the binding of high-affinity antibodies.

PubMed: 16597831
DOI: 10.1110/ps.052030506

実験手法

X-RAY DIFFRACTION (2.2 Å)