3KJ4

Structure of rat Nogo receptor bound to 1D9 antagonist antibody

3KJ4 の概要

• エントリーDOI: 10.2210/pdb3kj4/pdb
• 関連するPDBエントリー: 1OZN 1P8T
• 分子名称: Fab fragment 1D9 light chain, Fab fragment 1D9 heavy chain, Reticulon-4 receptor, ... (7 entities in total)
• 機能のキーワード: nogo receptor antagonist antibody, lrr, cell membrane, disulfide bond, glycoprotein, gpi-anchor, leucine-rich repeat, lipoprotein, membrane, receptor, immune system
• 由来する生物種: Mus musculus (mouse); 詳細
• 細胞内の位置: Cell membrane; Lipid-anchor, GPI-anchor (By similarity): Q99M75
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 160753.83

構造登録者

Silvian, L.F. (登録日: 2009-11-02, 公開日: 2010-09-22, 最終更新日: 2024-11-06)

主引用文献

Weinreb, P.H.,Wen, D.,Qian, F.,Wildes, C.P.,Garber, E.A.,Walus, L.,Jung, M.Y.,Wang, J.,Relton, J.K.,Amatucci, J.,Wang, R.,Porreca, F.,Silvian, L.,Meier, W.,Pepinsky, R.B.,Lee, D.H.
Resolution of disulfide heterogeneity in Nogo receptor 1 fusion proteins by molecular engineering.
Biotechnol Appl Biochem, 57:31-45, 2010

PubMed Abstract: NgRI (Nogo-66 receptor) is part of a signalling complex that inhibits axon regeneration in the central nervous system. Truncated soluble versions of NgRI have been used successfully to promote axon regeneration in animal models of spinal-cord injury, raising interest in this protein as a potential therapeutic target. The LRR (leucine-rich repeat) regions in NgRI are flanked by N- and C-terminal disulfide-containing 'cap' domains (LRRNT and LRRCT respectively). In the present work we show that, although functionally active, the NgRI(310)-Fc fusion protein contains mislinked and heterogeneous disulfide patterns in the LRRCT domain, and we report the generation of a series of variant molecules specifically designed to prevent this heterogeneity. Using these variants we explored the effects of modifying the NgRI truncation site or the spacing between the NgRI and Fc domains, or replacing cysteines within the NgRI or IgG hinge regions. One variant, which incorporates replacements of Cys²⁶⁶ and Cys³⁰⁹ with alanine residues, completely eliminated disulfide scrambling while maintaining functional in vitro and in vivo efficacy. This modified NgRI-Fc molecule represents a significantly improved candidate for further pharmaceutical development, and may serve as a useful model for the optimization of other IgG fusion proteins made from LRR proteins.

PubMed: 20815818
DOI: 10.1042/BA20100061

実験手法

X-RAY DIFFRACTION (3.1 Å)