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Protein crystallization, data collection, and structure solution.

RG Rhys Grinter
FM Faye C. Morris
RD Rhys A. Dunstan
PL Pok Man Leung
AK Ashleigh Kropp
MB Matthew Belousoff
SG Sachith D. Gunasinghe
NS Nichollas E. Scott
SB Simone Beckham
AP Anton Y. Peleg
CG Chris Greening
JL Jian Li
EH Eva Heinz
TL Trevor Lithgow
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Purified BonA proteins were screened for crystallization conditions using commercially available screens (∼800 conditions). Crystals grew from drops containing BonA-27N in medium composed of 0.2 M zinc acetate, 0.1 M sodium acetate, and 20% PEG 3350 (pH 4.5), and the crystals were optimized from this starting condition. The crystals were cryoprotected by increasing PEG 3350 concentration to 30% and flash cooled in liquid N2. Diffraction data were collected at 100 K at the Australian Synchrotron on selenomethionine-labeled crystals and processed in the space group P3121 to 1.65 Å. Heavy atom sites were located, phases were obtained using single-wavelength anomalous dispersion (SAD), and the initial model was built using Autosol from the Phenix package (60). Eight heavy atom sites were located; four of these sites were selenium, and four of these sites were zinc. The BonA-N27 model was improved manually in Coot and refined using Phenix refine and Refmac (60,–62). Analysis of the BonA-27N crystal structure was performed using the Phenix and CCP4 packages, noncrystallographic interfaces were predicted using PISA (46, 60, 63).

Copyright and License information:  ©2021 Grinter et al.
This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

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