| Budget Amount *help |
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1992: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1991: ¥5,400,000 (Direct Cost: ¥5,400,000)
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| Research Abstract |
In order to accelerate to get the structure of the proteins by x-ray protein crystallography, a trial has been made to facilitate crystal growth by trimming the target protein appropriately. This trial is in accordance with calls for more independent 3-dimensional structure information.
The target proteins were:F1-ATPase from a thermophilic bacterium, its alpha3beta3 complex, rat DNA polymerase beta, Env Z protein, PhoB protein, Band 3 protein, CamR repressor, Asp racemase and flagellin. Those proteins were subjected to a limited proteolysis by a number of proteases to look for appropriately trimmed molecules. The trimmed molecules were then purified and subjected to crystallization experiment.
The results are: 1.In case of rat DNA polymerase beta, a large C-terminal fragment (representing 80 % of the molecule) was found to crystallize to a suitable form. The intact molecule resisted to crystallization. The crystals diffracted to at least to 2.5 A^^ﾟ resolution, and the data collection and derivative search is now in progress. This indicates that the trimmed molecule does form good crystals. 2.41 k fragment of flagellin formed crystals. The intact flagellin formed helical aggregates under most of crystallization conditions. Although crystals were formed, the conditions are to be refined to get better crystals. 3.Appropriate fragments of PhoB protein and CamR repressor were found. They are now subjected to purification.
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