期刊
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY
卷 68, 期 -, 页码 521-530出版社
INT UNION CRYSTALLOGRAPHY
DOI: 10.1107/S0907444912004490
关键词
sulfur phasing; SAXS analysis; long-wavelength X-rays; death receptor 6
资金
- US Department of Energy (DOE) [DE-AC02-05CH11231]
- Office of Science, Office of Basic Energy Sciences of the DOE [DE-AC02-05CH11231]
- National Natural Science Foundation of China [30870483, 31070660, 31021062, 81072449]
- Ministry of Science and Technology of China [2009DFB30310, 2009CB918803, 2011CB911103]
- CAS [YZ200839, KSCX2-EW-J-3]
- NIGMS [NIH U54 GM074946]
A subset of tumour necrosis factor receptor (TNFR) superfamily members contain death domains in their cytoplasmic tails. Death receptor 6 (DR6) is one such member and can trigger apoptosis upon the binding of a ligand by its cysteine-rich domains (CRDs). The crystal structure of the ectodomain (amino acids 1348) of human death receptor 6 (DR6) encompassing the CRD region was phased using the anomalous signal from S atoms. In order to explore the feasibility of S-SAD phasing at longer wavelengths (beyond 2.5 angstrom), a comparative study was performed on data collected at wavelengths of 2.0 and 2.7 angstrom. In spite of sub-optimal experimental conditions, the 2.7 angstrom wavelength used for data collection showed potential for S-SAD phasing. The results showed that the Rano/Rp.i.m. ratio is a good indicator for monitoring the anomalous data quality when the anomalous signal is relatively strong, while d''/sig(d'') calculated by SHELXC is a more sensitive and stable indicator applicable for grading a wider range of anomalous data qualities. The use of the `parameter-space screening method' for S-SAD phasing resulted in solutions for data sets that failed during manual attempts. SAXS measurements on the ectodomain suggested that a dimer defines the minimal physical unit of an unliganded DR6 molecule in solution.
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