8C27

Domain 3 of Group B Streptococcus pilus protein as scaffold for the display of foreign epitopes

8C27 の概要

• エントリーDOI: 10.2210/pdb8c27/pdb
• 分子名称: PI-2a backbone protein, 1,2-ETHANEDIOL (3 entities in total)
• 機能のキーワード: pilus protein, scaffold, membrane protein, epitopes.
• 由来する生物種: Streptococcus agalactiae
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 29456.70

構造登録者

Veggi, D.,Cappelli, L.,Cozzi, R. (登録日: 2022-12-21, 公開日: 2024-01-10, 最終更新日: 2024-07-24)

主引用文献

Cappelli, L.,Cinelli, P.,Perrotta, A.,Veggi, D.,Audagnotto, M.,Tuscano, G.,Pansegrau, W.,Bartolini, E.,Rinaudo, D.,Cozzi, R.
Computational structure-based approach to study chimeric antigens using a new protein scaffold displaying foreign epitopes.
Faseb J., 38:e23326-e23326, 2024

PubMed Abstract: The identification and recombinant production of functional antigens and/or epitopes of pathogens represent a crucial step for the development of an effective protein-based vaccine. Many vaccine targets are outer membrane proteins anchored into the lipidic bilayer through an extended hydrophobic portion making their recombinant production challenging. Moreover, only the extracellular loops, and not the hydrophobic regions, are naturally exposed to the immune system. In this work, the Domain 3 (D3) from Group B Streptococcus (GBS) pilus 2a backbone protein has been identified and engineered to be used as a scaffold for the display of extracellular loops of two Neisseria gonorrhoeae membrane proteins (PorB.1b and OpaB). A computational structure-based approach has been applied to the design of both the scaffold and the model antigens. Once identified the best D3 engineerable site, several different chimeric D3 displaying PorB.1b and OpaB extracellular loops were produced as soluble proteins. Each molecule has been characterized in terms of solubility, stability, and ability to correctly display the foreign epitope. This antigen dissection strategy allowed the identification of most immunogenic extracellular loops of both PorB.1b and OpaB gonococcal antigens. The crystal structure of chimeric D3 displaying PorB.1b immunodominant loop has been obtained confirming that the engineerization did not alter the predicted native structure of this epitope. Taken together, the reported data suggest that D3 is a novel protein scaffold for epitope insertion and display, and a valid alternative to the production of whole membrane protein antigens. Finally, this work describes a generalized computational structure-based approach for the identification, design, and dissection of epitopes in target antigens through chimeric proteins.

PubMed: 38019196
DOI: 10.1096/fj.202202130R

実験手法

X-RAY DIFFRACTION (2.63 Å)