Abstract
The PsbO protein of photosystem II stabilizes the active-site manganese cluster and is thought to act as a proton antenna. To enable neutron diffraction studies, crystals of the β-barrel core of PsbO were grown in capillaries. The crystals were optimized by screening additives in a counter-diffusion setup in which the protein and reservoir solutions were separated by a 1% agarose plug. Crystals were cross-linked with glutaraldehyde. Initial neutron diffraction data were collected from a 0.25 mm3 crystal at room temperature using the MaNDi single-crystal diffractometer at the Spallation Neutron Source, Oak Ridge National Laboratory.Crystals of PsbO were grown in capillaries using an agarose-plug setup and were cross-linked using glutaraldehyde. Initial neutron diffraction data were collected to 2.3 Å resolution on the MaNDi instrument at the Spallation Neutron Source.
| Original language | English |
|---|---|
| Pages (from-to) | 525-531 |
| Number of pages | 7 |
| Journal | Acta Crystallographica Section F:Structural Biology Communications |
| Volume | 73 |
| Issue number | 9 |
| DOIs | |
| State | Published - Sep 2017 |
Funding
The authors thank the Deutsche Forschungsgemeinschaft (DFG) for financial support to the Collaborative Research Centre on ‘Protonation Dynamics in Protein Function’ (SFB1078, projects A5-Zouni/Dobbek and A4-Dau). The research at ORNL’s Spallation Neutron Source was sponsored by the Scientific User Facilities Division, Office of Basic Energy Sciences, US Department of Energy. The Office of Biological and Environmental Research supported research at Oak Ridge National Laboratory’s Center for Structural Molecular Biology (CSMB), using facilities supported by the Scientific User Facilities Division, Office of Basic Energy Sciences, US Department of Energy. For crystal screening, we acknowledge access to beamline P11 at PETRA III, DESY and beamlines 14.1 and 14.3 of the BESSY II storage ring (Berlin, Germany) via the Joint Berlin MX-Laboratory sponsored by the Helmholtz Zentrum Berlin für Materialien und Energie, the Freie Universität Berlin, the Humboldt-Universität zu Berlin, the Max-Delbrück-Centrum and the Leibniz-Institut für Molekulare Pharmakologie. We thank the members of the MX group at HZB and the P11 group at PETRA III for technical support. We would like to thank Rana Ali, Ruchira Chatterjee, Mohamed Ibrahim and Jan Kern for crystal handling and assistance during data collection and Rainer Dietrich for milling a Perspex capillary holder for room-temperature shipping.
| Funders | Funder number |
|---|---|
| Oak Ridge National Laboratory | |
| Office of Basic Energy Sciences | |
| Scientific User Facilities Division | |
| US Department of Energy | |
| Basic Energy Sciences | |
| Oak Ridge National Laboratory | |
| Canadian Society for Molecular Biosciences | |
| Deutsche Forschungsgemeinschaft | A4-Dau, SFB1078 |
Keywords
- PsbO
- agarose
- capillary
- counter-diffusion
- cross-linking
- neutron diffraction
- photosystem II
