Acta Crystallographica Section D

Biological Crystallography

Volume 61, Part 11 (November 2005)

research papers

Acta Cryst. (2005). D61, 1533-1540 [ doi:10.1107/S0907444905028416 ]

Comparison of phasing methods for sulfur-SAD using in-house chromium radiation: case studies for standard proteins and a 69 kDa protein

N. Watanabe, Y. Kitago, I. Tanaka, J. Wang, Y. Gu, C. Zheng and H. Fan

Abstract: Phasing of the crystal structures of four standard proteins (lysozyme, trypsin, glucose isomerase and thaumatin) and a novel 69 kDa protein from Thermus thermophilus, TT0570, was performed using the single-wavelength anomalous diffraction of S atoms intrinsically present within the native protein molecules. To utilize the sulfur anomalous diffraction, the data sets were collected using the loopless data-collection method with chromium K [alpha] X-rays of wavelength 2.29 Å. Three phasing methods, MLPHARE, SHARP and OASIS-2004, were tested in combination with the DM or SOLOMON density-modification method. The results showed that the solvent contents are still an important factor for phasing with the S-SAD method, even when longer wavelength Cr K [alpha] radiation is used. Of the three procedures, the improved direct phasing of OASIS-2004 with its implemented fragment feedback to the direct-method probability calculation gave the best results in determining the initial phases. For all five proteins, almost the entire models could be built automatically.

Keywords: single-wavelength anomalous diffraction; sulfur SAD; chromium X-ray radiation.