The capability to reach wavelengths of up to 3.1 A ̊ 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 = 2.69 A ̊ (4.6 keV) allowed the crystal structure determination by sulfur SAD phasing of Cdc23Nterm, a subunit of the multimeric anaphase-promoting complex (APC/C). At this energy, Cdc23Nterm has an expected Bijvoet ratio h|Fanom|i/hFi 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 mm in diameter allowed crystal twinning to be overcome. The crystals diffracted to 3.1 A ̊ resolution, with unit-cell parameters a = b = 61.2, c = 151.5 A ̊ , and belonged to space group P43. The refined structure to 3.1 A ̊ 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.
Data collection with a tailored X-ray beam size at 2.69 Å wavelength (4.6 keV): Sulfur SAD phasing of Cdc23Nterm / Cianci, Michele; Groves, Matthew R.; Barford, David; Schneider, Thomas R.. - In: ACTA CRYSTALLOGRAPHICA. SECTION D, BIOLOGICAL CRYSTALLOGRAPHY. - ISSN 0907-4449. - ELETTRONICO. - 72:3(2016), pp. 403-412. [10.1107/S2059798315010268]
Data collection with a tailored X-ray beam size at 2.69 Å wavelength (4.6 keV): Sulfur SAD phasing of Cdc23Nterm
CIANCI, MICHELE
;
2016-01-01
Abstract
The capability to reach wavelengths of up to 3.1 A ̊ 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 = 2.69 A ̊ (4.6 keV) allowed the crystal structure determination by sulfur SAD phasing of Cdc23Nterm, a subunit of the multimeric anaphase-promoting complex (APC/C). At this energy, Cdc23Nterm has an expected Bijvoet ratio h|Fanom|i/hFi 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 mm in diameter allowed crystal twinning to be overcome. The crystals diffracted to 3.1 A ̊ resolution, with unit-cell parameters a = b = 61.2, c = 151.5 A ̊ , and belonged to space group P43. The refined structure to 3.1 A ̊ 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.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.