Extended Data Fig. 2: Alignment of P. falciparum FIKK protein sequences allows for accurate determination of the FIKK kinase domain starting amino acid.

a, Amino acid sequence conservation in P. falciparum FIKKs was assessed using the PRALINE Multiple Sequence Alignment Software¹¹⁶. Conservation values reflect the normalised average of BLOSUM62 scores for each alignment column and range from 0 (low conservation) to 10 (high conservation). Sequence position is with respect to P. falciparum FIKK8 as the reference sequence. Green shading illustrates the FIKK kinase domain. The eponymous F-I-K-K motif is represented in red. b, Alignment of all FIKK sequences from P. falciparum including pseudokinases FIKK7.2 and FIKK14 using the T-Coffee multiple sequence alignment program¹¹⁷ available in the Jalview software¹¹⁸. Shown is the alignment for the FIKK kinase domain. FIKK4.2 insertion (residues 381-953) has been removed to simplify visualisation. Encircled in red are the amino acids chosen as a starting point for recombinant expression of P. falciparum FIKK kinase domains. The ClustalX colour scheme was used to assign colour to amino acids with the following criteria: Blue – Hydrophobic (A, I, L, M, F, W, V, C); Red – Positively charged (K, R); Magenta – Negatively charged (D, E); Green – Polar (N, Q, S, T); Orange – Glycine (G); Yellow – Proline (P); Cyan – Aromatic (H, Y); White – unconserved amino acids. Below the alignment is indicated the conservation score which measures the number of physicochemical properties conserved for each column of the alignment. Its calculation is based on¹¹⁹. Conserved columns are indicated by * (score of 11), conservation score then ranges between 10 (high conservation) and 0 (no conservation). Hyphens denote gaps.