LiBRC1 is a more potent binder of LiRAD51 than LiBRC2.
(A) Sequence alignment of the two L. infantum BRC repeats with HsBRC4 and point mutants LiBRC1.1 and LiBRC2.1. Competition FP binding results are shown on the right. SE, standard error of fit. (B) Coomassie stained SDS–PAGE gel analysis of L. infantum BRC repeat affinity pull-down of LiRAD51ATPase (residues 134–386). (C) Direct FP titration of LiRAD51ATPase into fluorescein-tagged LiBRC1 (5 nM). Data shown are the means of triplicate measurements ±SE. (D) Competition FP titrations of GB1-fused LiBRC1 and LiBRC2. Fluorescently labelled LiBRC1 probe (5 nM) was pre-incubated with 500 nM LiRAD51ATPase, to which GB1-LiBRC dilution series were added. Data shown are the means of triplicate measurements ±SD.