首页   按字顺浏览 期刊浏览 卷期浏览 The application of direct methods and Patterson interpretation to high‐resolution nativ...
The application of direct methods and Patterson interpretation to high‐resolution native protein data

 

作者: G. M. Sheldrick,   Z. Dauter,   K. S. Wilson,   H. Hope,   L. C. Sieker,  

 

期刊: Acta Crystallographica Section D  (WILEY Available online 1993)
卷期: Volume 49, issue 1  

页码: 18-23

 

ISSN:1399-0047

 

年代: 1993

 

DOI:10.1107/S0907444992007364

 

出版商: International Union of Crystallography

 

数据来源: WILEY

 

摘要:

Conventional small‐molecule methods of solving the phase problem from native data alone, without the use of heavy‐atom derivatives, known fragment geometries or anomalous dispersion, have been tested on 0.9 Å resolution data for two small proteins: rubredoxin, fromDesulfovibrio vulgaris, and crambin. The presence of three disulfide bridges in crambin and an FeS4unit in rubredoxin enabled automated Patterson interpretation as well as direct methods to be tried. Although both structures were already well established, the known structures were not used in the phasing attempts, except for identifying successful solutions. Direct methods were not successful for crambin, although the correct phases were stable to phase refinement and gave figures of merit clearly superior to any obtained in theca500 000 random starting phase sets that were refined. It appears that the presence of an iron atom in rubredoxin reduces the scale of the search problem by many orders of magnitude, but at the cost of producing `over‐consistent' phase sets that overemphasize the iron atom and involve partial loss of enantiomorph information. However, about 1% of direct‐methods trials were successful for rubredoxin, giving mean phase errors of about 56° (for allE>1.2) that could be reduced to about 20° by standardE‐Fourier recycling methods. Limiting the resolution of the data degraded the quality of the solutions and suggested that the limiting resolution for routine direct‐methods solution of rubredoxin is about 1.2 Å. With the 0.9 Å data, automated Patterson interpretation convincingly finds the three disulfide bridges in crambin and the FeS4unit in rubredoxin, and in both casesE‐Fourier recycling starting from these `heavier' atoms yields almost the complete structure. Whereas crambin could only be solved in this way at very high resolution, rubredoxin could be solved by Patterson interpretation down to 1.6 Å. These results emphasize the benefits of collecting protein data to the highest possible resolution, and indicate that when a few `heavier' atoms are present, it may prove possible in favorable cases to solve the phase problem from a single native data set col

 

点击下载:  PDF (661KB)



返 回