Abstract:
:A random conical tilt reconstruction of negatively stained Saccharomyces cerevisiae fatty acid synthase was used as a model to compute a three-dimensional reconstruction from untilted stain specimens of the molecules in multiple orientations using a three-dimensional projection alignment method. The resulting structure (24 A resolution) has a more uniform resolution than the initial structure and the handedness revealed in the random conical tilt method is preserved. In a similar approach, this model was used to compute a 21-A-resolution frozen-hydrated structure from untilted specimens of the molecules in multiple orientations. Even though the reconstructions are in close agreement, the stain structure appears to enhance the protein density associated with less robust features. These procedures significantly reduce the time and effort required to obtain a three-dimensional reconstruction from frozen-hydrated data with a resolution that is comparable to the best obtained by more laborious methods. The agreement between the stain and frozen-hydrated reconstructions affords convincing evidence concerning the validity of the structure and the information afforded by the two reconstructions significantly enhances the structural analysis of the molecule.
journal_name
J Struct Bioljournal_title
Journal of structural biologyauthors
Kolodziej SJ,Penczek PA,Stoops JKdoi
10.1006/jsbi.1997.3911subject
Has Abstractpub_date
1997-11-01 00:00:00pages
158-67issue
2eissn
1047-8477issn
1095-8657pii
S1047-8477(97)93911-3journal_volume
120pub_type
杂志文章abstract::The configuration of the active site of E2 ligases, central enzymes in the ubiquitin/ubiquitin-like protein (Ub/Ubl) conjugation systems, has long puzzled researchers. Taking advantage of the wealth of newly available structures and sequences of E2s from diverse organisms, we performed a large-scale comparative analys...
journal_title:Journal of structural biology
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journal_title:Journal of structural biology
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journal_title:Journal of structural biology
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journal_title:Journal of structural biology
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journal_title:Journal of structural biology
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journal_title:Journal of structural biology
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journal_title:Journal of structural biology
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journal_title:Journal of structural biology
pub_type: 杂志文章
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journal_title:Journal of structural biology
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journal_title:Journal of structural biology
pub_type: 杂志文章
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journal_title:Journal of structural biology
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journal_title:Journal of structural biology
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journal_title:Journal of structural biology
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journal_title:Journal of structural biology
pub_type: 杂志文章
doi:10.1016/j.jsb.2004.10.010
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journal_title:Journal of structural biology
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