Fig. 3: FIKK kinases evolved divergent substrate specificities conserved among Laverania species.

a, Extended Data Fig. 3 data represented as a heat map (i). The ³²P incorporation values were normalized to 20 (the number of possible natural amino acids) and are shown as log₂(x) where negative values (blue cells) indicate disfavoured amino acids and positive values (red cells) indicate favoured amino acids. A PWM logo generated with FIKK8 raw OPAL data (ii). PWMs depict the preference of the kinase for all 20 amino acids at every substrate position. For ease of visualization, the PWM logo displays amino acids with scores above an arbitrary threshold of 2.5 (Methods). Amino acid colours are set as follows: acidic negatively charged (D, E), red; basic positively charged (R, K, H), blue; polar uncharged (N, Q), purple; non-polar (A, I, L, M, F, V, P), black; phosphorylatable or special (S, T, Y, C, G), green. b, A heat map representation of OPAL data for basophilic FIKK1 (left), acidophilic FIKK9.1 (middle) and tyrosine kinase FIKK13 (right). PWM logos generated from raw OPAL data are displayed below the corresponding heat maps. OPAL membrane images are available in Supplementary Fig. 5. c, PWM logos generated with PgFIKKs raw OPAL data. See a(i) caption and Supplementary Fig. 6. d, A maximum-likelihood phylogenetic tree of Laverania FIKK sequences built using FIKK8 kinases and two avian malaria FIKKs (P. relictum FIKK PRELSG_0112400 and P. gallinaceum FIKK PGAL8A_00108200) as an outgroup. One hundred bootstrap replicates were generated to assess branch support⁸⁹. All orthologue clades have maximum branch support (one out of one). Branches between paralogues are highlighted in red if they are >0.5. The triangle length represents the divergence between FIKK sequences within a specific clade. The colour code identifies the kinases substrate specificities as follows: blue, basophilic; red, acidophilic; purple, tyrosine kinase; white, uncharacterized. Sequence logos for each clade are given for the P. falciparum kinase copy. The tree contains N = 131 sequences in total.