Journal
PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS
Volume 88, Issue 1, Pages 187-195Publisher
WILEY
DOI: 10.1002/prot.25786
Keywords
deep sequencing; high-affinity design; host-pathogen interactions; Plasmodium falciparum; Rosetta
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Funding
- Benoziyo Endowment Fund for the Advancement of Science
- European Research Council [815379]
- Wellcome [100993/Z/13/Z]
- Sir Henry Wellcome Postdoctoral Fellowship [107366/Z/15/Z]
- UK Medical Research Council [MR/K501281/1]
- Wellcome Trust [106917/Z/15/Z]
- Lister Institute
- Wellcome Trust [107366/Z/15/Z] Funding Source: Wellcome Trust
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Many human pathogens use host cell-surface receptors to attach and invade cells. Often, the host-pathogen interaction affinity is low, presenting opportunities to block invasion using a soluble, high-affinity mimic of the host protein. The Plasmodium falciparum reticulocyte-binding protein homolog 5 (RH5) provides an exciting candidate for mimicry: it is highly conserved and its moderate affinity binding to the human receptor basigin (K-D >= 1 mu M) is an essential step in erythrocyte invasion by this malaria parasite. We used deep mutational scanning of a soluble fragment of human basigin to systematically characterize point mutations that enhance basigin affinity for RH5 and then used Rosetta to design a variant within the sequence space of affinity-enhancing mutations. The resulting seven-mutation design exhibited 1900-fold higher affinity (K-D approximately 1 nM) for RH5 with a very slow binding off rate (0.23 h(-1)) and reduced the effective Plasmodium growth-inhibitory concentration by at least 10-fold compared to human basigin. The design provides a favorable starting point for engineering on-rate improvements that are likely to be essential to reach therapeutically effective growth inhibition.
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