Challenges of sulfur SAD phasing as a routine method in macromolecular crystallography

DOI: 10.1107/S0909049511049004
PMID: 22186640

Authors: James Doutch (University of Liverpool) , Mike Hough (University of Essex) , Samar Hasnain (University of Liverpool) , Richard Strange (University of Liverpool)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Journal Of Synchrotron Radiation , VOL 19 (1) , PAGES 19-29

State: Published (Approved)
Published: December 2011

Abstract: The sulfur SAD phasing method allows the determination of protein structures de novo without reference to derivatives such as Se-methionine. The feasibility for routine automated sulfur SAD phasing using a number of current protein crystallography beamlines at several synchrotrons was examined using crystals of trimeric Achromobacter cycloclastes nitrite reductase (AcNiR), which contains a near average proportion of sulfur-containing residues and two Cu atoms per subunit. Experiments using X-ray wavelengths in the range 1.9-2.4 Å show that we are not yet at the level where sulfur SAD is routinely successful for automated structure solution and model building using existing beamlines and current software tools. On the other hand, experiments using the shortest X-ray wavelengths available on existing beamlines could be routinely exploited to solve and produce unbiased structural models using the similarly weak anomalous scattering signals from the intrinsic metal atoms in proteins. The comparison of long-wavelength phasing (the Bijvoet ratio for nine S atoms and two Cu atoms is ~1.25% at ~2 Å) and copper phasing (the Bijvoet ratio for two Cu atoms is 0.81% at ~0.75 Å) for AcNiR suggests that lower data multiplicity than is currently required for success should in general be possible for sulfur phasing if appropriate improvements to beamlines and data collection strategies can be implemented.

Journal Keywords: Single-Wavelength Anomalous Diffraction; Automated S-Sad; Data Redundancy; Cu-Sad; Phasing; Radiation Damage.

Subject Areas: Biology and Bio-materials


Instruments: I02-Macromolecular Crystallography , I03-Macromolecular Crystallography

Other Facilities: SOLEIL