Fig. 4: Experimental characterization and modeling of calcium-activated hEndoU in complex with RNA.
From: Molecular basis for the calcium-dependent activation of the ribonuclease EndoU
A Combined 1H-15N chemical shift perturbations between calcium-activated 13C15N-hEndoU and in complex with 2’-F RNA. Source data are provided as a Source Data file. B RNA binding interface mapping on calcium-activated hEndoU. C Electrostatic surface potential of calcium-activated hEndoU. D Structure alignment of the ensemble of models (N = 14) for calcium-activated hEndoU in complex with RNA. The alignment was performed on the protein region. E Detailed view of a representative model of the ensemble with highlight on protein-RNA interactions. A close-up view of interactions at U2-U3 is also shown. F RNA binding to calcium-activated hEndoU mutants monitored by EMSA. The regions corresponding to the free Cy5-labeled RNA, the low molecular weight complex (LMW), and the higher molecular weight complexes (HMW), are highlighted. Experiments were done in triplicate. For each experiment, the relative fraction of Free RNA, LMW, and HMW complexes were computed for each replicate, and the resulting means (plotted values) and standard deviations (error bars) were plotted at 10 eq. hEndoU.
