High Throughput Screens Yield Small Molecule Inhibitors of Leishmania CRK3:CYC6 Cyclin-Dependent Kinase
Figure 1
Expression and purification of the Leishmania CRK3:CYC6.
(A) CRK3his and CYC6his were expressed in E. coli individually (lanes 1 and 2 respectively). Coomassie-stained SDS-PAGE. (B) Co-expressed CRK3his and CYC6his purified by Nickel-chelate and gel filtration chromatography. Coomassie-stained SDS-PAGE. (C) Co-expression of CRK3 and CYC6his and purification of the CRK3:CYC6his by Nickel-chelate and ion-exchange chromatography. Coomassie-stained SDS-PAGE. (D) Activation of CRK3his histone H1 kinase in the presence of increasing concentrations of CYC6his. All lanes contain 1.25 µg of CRK3 and 0.025 µg, 0.05 µg, 0.075 µg, 0.1 µg of CYC6 (lanes 2–5).