De novo crystal structure determination of double stranded RNA binding domain using only the sulfur anomalous diffraction in SAD phasing.

ABSTRACT: Single-wavelength anomalous dispersion (SAD)-phasing using sulfur as the unique anomalous scatterer is a powerful method to solve the phase problem in protein crystallography. However, it is not yet widely used by non-expert crystallographers. We report here the structure determination of the double stranded RNA binding domain of human dihydrouridine synthase using the sulfur-SAD method and highly redundant data collected at 1.8 ​Å ("off-edge"), at which the estimated overall anomalous signal was 1.08%. High multiplicity data were collected on a single crystal rotated along the ϕ or ω axis at different κ angles, with the primary beam intensity being attenuated from 50% to 95%, compared to data collection at 0.98 ​Å, to reduce radiation damage. SHELXD succeeded to locate 14 out 15 su

SUBMITTER: Guimaraes BG 

PROVIDER: S-EPMC8244422 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature