Extended Data Fig. 3: FIKK8 OPAL membrane.
From: The fast-evolving FIKK kinase family of Plasmodium falciparum can be inhibited by a single compound
An OPAL membrane constituted of 9-mer peptides with the general sequences A-X-X-X-X-S-X-X-X-X-A (top panel), A-X-X-X-X-T-X-X-X-X-A (middle panel) or A-X-X-X-X-Y-X-X-X-X-A (bottom panel) was used to assess FIKK8 preferred phosphorylation motif. X represents any natural amino acid except for S, T, Y or C. For each peptide, one of the 20 naturally occurring amino acids is fixed at each one of the 8 positions surrounding the phosphorylatable residue (S, T or Y). The membrane was incubated in the presence of recombinant FIKK8 kinase domain and [ɣ-32P]-ATP. After several washes, the membrane was exposed overnight to a phosphorscreen. The radioactivity incorporated into each peptide was determined by scanning the phosphorscreen with a phosphorimager giving the radiograph visible in this figure. Plotting the intensity pattern of the array enables the identification of preferred phosphorylation motifs (Fig. 3a(ii)) and reveals amino acids that are less favoured in a peptide sequence.
