Table 2. Predicted functional characteristics of PfCyRPA SNPs.

Individual FASTA files of PfCyRPA alleles were threaded through the crystal structure and the impact of the mutant versions of the protein was evaluated for predicted binding affinity of PfCyRPA to its binding partner PfRH5 or neutralizing human mAbs. The positions of the SNPs were determined using structural data from Chen et al., 2017. The binding energy alternation from the SNPs was predicted by FoldX version 5.0. Predicted binding energies are shown for reference and mutant alleles of the protein in Kcal/Mol for each SNP. Changes between the two are shown as ΔΔG (Kcal/mol). A negative ΔΔG indicates a predicted increase in PfCyRPA stability while a positive ΔΔG is associated with a predicted decrease in PfCyRPA stability.

SNP	Blade	Predicted impact on interaction with RH5 and antibodies Cy.003, Cy.004, and Cy.007	ΔΔG of CyRPA stability (Kcal/mol)
R31H	6	Minor effect	1.91
T37A	6	Minor effect	0.45
F41L	6	Minor effect	1.23
R50C	1	Minor effect on antibody binding, but may improve RH5 binding by removing repulsion between CyRPA R50 and RH5 K504	1.11
D110N	2	Minor effect	−2.02
I114V	2	Minor effect	0.57
V165I	3	Minor effect; may alter CyRPA structure	−0.99
F187L	3	Minor effect on antibody binding, but may affect binding to RH5	0.2
I196F	3	Minor effect on antibody binding, may affect binding to RH5; may alter CyRPA structure	9.33
K211Q	4	Minor effect	0.95
D236N	4	Add a new N-glycosylation site, minor effect	0.05
D236V	4	Alter local structure through steric clash with S233, minor effect on antibody and RH5 binding	0.09
N270T	4	Minor effect on antibody and RH5 binding; alter CyRPA structure, but abolishes a hydrogen bond between CyRPA N218 and N270	−0.03
V292F	5	Minor effect; may alter CyRPA structure	8.54
N338D	6	Minor effect	1.75