5FTP

sulfur SAD phasing of Cdc23Nterm: data collection with a tailored X- ray beam size at 2.69 A wavelength (4.6 keV)

Summary for 5FTP

• Entry DOI: 10.2210/pdb5ftp/pdb
• Descriptor: ANAPHASE-PROMOTING COMPLEX SUBUNIT 8 (2 entities in total)
• Functional Keywords: sad phasing, sulfur, soft x-rays, long wavelength, low resolution, cell cycle
• Biological source: SCHIZOSACCHAROMYCES POMBE
• Total number of polymer chains: 2
• Total formula weight: 67738.86

Authors

Cianci, M.,Groves, M.R.,Barford, D.,Schneider, T.R. (deposition date: 2016-01-14, release date: 2016-03-16, Last modification date: 2024-05-08)

Primary citation

Cianci, M.,Groves, M.R.,Barford, D.,Schneider, T.R.
Data Collection with a Tailored X-Ray Beam Size at 2.69 A Wavelength (4.6 Kev): Sulfur Sad Phasing of Cdc23Nterm
Acta Crystallogr.,Sect.D, 72:403-, 2016

PubMed Abstract: The capability to reach wavelengths of up to 3.1 Å at the newly established EMBL P13 beamline at PETRA III, the new third-generation synchrotron at DESY in Hamburg, provides the opportunity to explore very long wavelengths to harness the sulfur anomalous signal for phase determination. Data collection at λ = 2.69 Å (4.6 keV) allowed the crystal structure determination by sulfur SAD phasing of Cdc23(Nterm), a subunit of the multimeric anaphase-promoting complex (APC/C). At this energy, Cdc23(Nterm) has an expected Bijvoet ratio〈|Fanom|〉/〈F〉of 2.2%, with 282 residues, including six cysteines and five methionine residues, and two molecules in the asymmetric unit (65.4 kDa; 12 Cys and ten Met residues). Selectively illuminating two separate portions of the same crystal with an X-ray beam of 50 µm in diameter allowed crystal twinning to be overcome. The crystals diffracted to 3.1 Å resolution, with unit-cell parameters a = b = 61.2, c = 151.5 Å, and belonged to space group P43. The refined structure to 3.1 Å resolution has an R factor of 18.7% and an Rfree of 25.9%. This paper reports the structure solution, related methods and a discussion of the instrumentation.

PubMed: 26960127
DOI: 10.1107/S2059798315010268

Experimental method

X-RAY DIFFRACTION (3.1 Å)