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. Author manuscript; available in PMC: 2016 Jan 5.
Published in final edited form as: Nature. 2015 Jun 18;522(7556):315–320. doi: 10.1038/nature14451

Figure 4. eFF2.

Figure 4

a. eEF2 promotes the GTP-dependent translocation of the ribosome along mRNA during protein synthesis.

b. Homology model of Plasmodium falciparum eEF2. The mapped mutations from each strain are colour coded by EC50 fold (red high, amber moderate, green low).

c. Live cell imaging of P. falciparum expressing an extra copy of eEF2 (WT) fused to GFP. The image is representative of >50 parasites visualized on two independent occasions.

d. Protein and DNA/RNA synthesis were evaluated by measuring the incorporation of [35S]-labelled methionine and cysteine ([35S]-Met/Cys) (upper panel) and [3H]-labelled hypoxanthine (lower panel) into asynchronous 3D7 wild-type (○) and 3D7 DDD107498-resistant line (eEF2-E134A/P754A) (●) after 40 min incubation with DDD107498, cycloheximide or actinomycin D. Radiolabeled incorporation, measured as cpm, was normalised as % of incorporation against inhibitor concentration (means ± s.d.; n=3 independent experiments each run in duplicate).

e. The EC50 values for transfectants against DDD107498 (means ± s.d.; n=4-7 independent experiments, each run in duplicate). Statistical significance was determined by the Mann-Whitney U test: *P<0.05; **P<0.01.

f. DDD107498-resistant line (eEF2-Y186N) transfected episomally with plasmids expressing either WT-eEF2 or eEF2-Y186N (means ± s.d.; n=3 independent experiments each run in duplicate).