5MI0

A thermally stabilised version of Plasmodium falciparum RH5

5MI0 の概要

• エントリーDOI: 10.2210/pdb5mi0/pdb
• 分子名称: Reticulocyte binding-like protein 5,Reticulocyte binding protein 5, MONOCLONAL ANTIBODY 9AD4, ... (4 entities in total)
• 機能のキーワード: malaria immunogen design rosetta pross, immune system
• 由来する生物種: Plasmodium falciparum; 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 97742.87

構造登録者

Campeotto, I.,Goldenzweig, A.,Davey, J.,Barfod, L.,Marshall, J.M.,Silk, S.E.,Wright, K.E.,Draper, S.J.,Higgins, M.K.,Fleishman, S.J. (登録日: 2016-11-27, 公開日: 2016-12-28, 最終更新日: 2024-10-16)

主引用文献

Campeotto, I.,Goldenzweig, A.,Davey, J.,Barfod, L.,Marshall, J.M.,Silk, S.E.,Wright, K.E.,Draper, S.J.,Higgins, M.K.,Fleishman, S.J.
One-step design of a stable variant of the malaria invasion protein RH5 for use as a vaccine immunogen.
Proc. Natl. Acad. Sci. U.S.A., 114:998-1002, 2017

PubMed Abstract: Many promising vaccine candidates from pathogenic viruses, bacteria, and parasites are unstable and cannot be produced cheaply for clinical use. For instance, Plasmodium falciparum reticulocyte-binding protein homolog 5 (PfRH5) is essential for erythrocyte invasion, is highly conserved among field isolates, and elicits antibodies that neutralize in vitro and protect in an animal model, making it a leading malaria vaccine candidate. However, functional RH5 is only expressible in eukaryotic systems and exhibits moderate temperature tolerance, limiting its usefulness in hot and low-income countries where malaria prevails. Current approaches to immunogen stabilization involve iterative application of rational or semirational design, random mutagenesis, and biochemical characterization. Typically, each round of optimization yields minor improvement in stability, and multiple rounds are required. In contrast, we developed a one-step design strategy using phylogenetic analysis and Rosetta atomistic calculations to design PfRH5 variants with improved packing and surface polarity. To demonstrate the robustness of this approach, we tested three PfRH5 designs, all of which showed improved stability relative to wild type. The best, bearing 18 mutations relative to PfRH5, expressed in a folded form in bacteria at >1 mg of protein per L of culture, and had 10-15 °C higher thermal tolerance than wild type, while also retaining ligand binding and immunogenic properties indistinguishable from wild type, proving its value as an immunogen for a future generation of vaccines against the malaria blood stage. We envision that this efficient computational stability design methodology will also be used to enhance the biophysical properties of other recalcitrant vaccine candidates from emerging pathogens.

PubMed: 28096331
DOI: 10.1073/pnas.1616903114

実験手法

X-RAY DIFFRACTION (2.35 Å)