SR INSTRUMENTATION FOR OPTIMIZED ANOMALOUS SCATTERING AND HIGH-RESOLUTION STRUCTURE STUDIES OF PROTEINS AND NUCLEIC-ACIDS

被引:15
作者
DEACON, A
HABASH, J
HARROP, SJ
HELLIWELL, JR
HUNTER, WN
LEONARD, GA
PETERSON, M
HADENER, A
KALB, AJ
ALLINSON, NM
CASTELLI, C
MOON, K
MCSWEENEY, S
GONZALEZ, A
THOMPSON, AW
EALICK, S
SZEBENYI, DM
WALTER, R
机构
[1] UNIV MANCHESTER,DEPT CHEM,MANCHESTER M13 9PL,LANCS,ENGLAND
[2] UNIV BASEL,BASEL,SWITZERLAND
[3] WEIZMANN INST SCI,IL-76100 REHOVOT,ISRAEL
[4] UNIV YORK,DEPT ELECTR,YORK YO1 5DD,N YORKSHIRE,ENGLAND
[5] SERC,DARESBURY LAB,DRAL,WARRINGTON WA4 4AD,CHESHIRE,ENGLAND
[6] EUROPEAN MOLEC BIOL LAB,EUROPEAN SYNCHROTRON RADIAT FACIL,F-38042 GRENOBLE,FRANCE
[7] CORNELL UNIV,MACCHESS,CHESS,ITHACA,NY
关键词
D O I
10.1063/1.1145956
中图分类号
TH7 [仪器、仪表];
学科分类号
0804 ; 080401 ; 081102 ;
摘要
Crystal structure solution by anomalous dispersion methods has been greatly facilitated using the rapidly tunable station 9.5 at the Daresbury SRS. Both SIROAS and MAD techniques, with IP data, have been used in the phasing of a brominated nucleotide and a seleno deaminase, respectively. The electron density maps in each case are interpretable. Throughput of projects could be improved upon with a better duty cycle detector. Another category of data collection is that at very high resolution. Detailed structure refinement pushes the limits of resolution and data quality. Station 9.5 has been used to collect high resolution (1.4 Å) native data for the protein concanavalin A. This utilized very short wavelengths (0.7 Å), the image plate, and crystal freezing. A total of 155 407 measurements from two crystals benefited from the on-line nature of the IP detector device, but a slow and quick pass are required to capture the full dynamic range of the data. There are data seen to 1.2 Å and beyond for a pure Mn substituted form of the protein, but a higher intensity still is required to actually record these data. By comparison, trials at CHESS, on a multipole wiggler (station A1) with a CCD (without image intensifier) system, yield native concanavalin A data to 0.98 Å and beyond. This demonstrates that the combination of yet higher intensity and the ease of use of a CCD offers worthwhile improvements; in this case an increase in the data by a factor of (1.4/0.98)3, thus at least doubling the data to parameter ratio for protein structure model refinement and potentially opening up direct structure determination of proteins of the size of concanavalin A (25 kDa). Finally, possibilities at ESRF and further detector developments, such as mosaic CCDs and scintillator coatings, offer further impetus for the field. These include more intense rapidly tunable beams for anomalous dispersion-based structure solution and ''ideal'' higher resolution data collection and reactivity studies. ESRF BL19 is described; facilities on BL19 will include a system for freezing and storing crystals at cryogenic temperatures, so that data can be recorded from the same crystal on different runs. Overall, there have been tremendous strides made in this field in the last 15 years, and yet further improvements are to come. © 1995 American Institute of Physics.
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收藏
页码:1287 / 1292
页数:6
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