Characterization and preliminary attempts for derivatization of crystals of large ribosomal subunits from Haloarcula marismortui diffracting to 3 Å resolution
An improved form of crystals of large (50 S) ribosomal subunits from Haloarcula marismortui, formally named Halobacterium marismortui, diffracting to 3 Å resolution, has been obtained by the addition of 1 m m-Cd 2+ to the crystallization medium, which contained more than 1·9 m of other salts. The im...
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Veröffentlicht in: | Journal of molecular biology 1991-11, Vol.222 (1), p.11-15 |
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Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | An improved form of crystals of large (50 S) ribosomal subunits from
Haloarcula marismortui, formally named
Halobacterium marismortui, diffracting to 3 Å resolution, has been obtained by the addition of 1 m
m-Cd
2+ to the crystallization medium, which contained more than 1·9
m of other salts. The improved crystals, grown from functionally active particles to an average size of 0·3 mm × 0·3 mm × 0·08 mm, are isomorphous with the previously reported ones, which diffracted to 4·5 Å. They are of space group
C222
1, cell dimensions
a = 210
A
̊
, b = 300
A
̊
, c = 581
A
̊
, and contain one particle in the asymmetric unit. Their superior internal order is reflected not only in their high resolution, but also in their reasonable mosaicity (less than 0·3 °). In contrast to the previously grown crystals, the new ones are of adequate mechanical strength and survive well the shock-cooling treatment. Due to their weak diffracting power, all crystallographic studies have been performed with synchrotron radiation. At cryotemperature, these crystals showed no measurable decay for a few days of irradiation and a complete diffraction data set could be collected from a single crystal. Efforts for initial phasing by specific and quantitative derivatization with superdense heavy-atom clusters are in progress. |
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ISSN: | 0022-2836 1089-8638 |
DOI: | 10.1016/0022-2836(91)90730-T |