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SAD experiments were recorded at beamline X06SA/PXI (SLS, Villingen) and native datasets at beamline ID23-1 (ESRF, Grenoble).
Native datasets were collected at APS LS-CAT and processed using the software HKL2000.
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A native dataset was collected at NSLS beamline X6A on a crystal that diffracted to 2.2 Å resolution.
The final resolution was extended to 1.7 Å using a high resolution native dataset and refined without NCS.
A native dataset was collected to a resolution of 1.5 Å at ESRF beam line (Grenoble, France) BM14.
Following initial rebuilding using the SeMet data, refinement was switched to minimize against the isomorphous native dataset.
Native dataset and a complete MAD dataset from single crystals of SeMet incorporated NBD94566 663 were collected at 100 K on beamline BL26B2 at SPring-8 using a Mar CCD 225 detector.
The quality and cryo-protection of the SeMet crystals was improved considerably and a peak-wavelength SAD dataset of better resolution (2.7 Å) was collected at the European Synchrotron Radiation Facility ESRFF, beamline BM14) along with a native dataset (2.4 Å).
The refinement was carried out with CNsimulatedted annealing [36] and Refmac [37] alternately using the 2.0 Å methylated native dataset and resulted in Rwork value of 20.30% and Rfree value of 25.14% (Table 2).
For the native dataset, significant population structure was observed between Florida and all other Caribbean populations (FST = 0.117±0.011), except Panama (FST = 0.042); however, after sequential Bonferroni correction, none of the comparisons remained significant (Table 4).
The crystals were cryoprotected in a buffer of 100 mM sodium acetate, pH 4.0, containing 2.5 M NaCl, and 35% glycerol, and the native dataset used to determine the structure was collected at APS beamline ID-24E.
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