Document Detail


Capture and visualization of a catalytic RNA enzyme-product complex using crystal lattice trapping and X-ray holographic reconstruction.
MedLine Citation:
PMID:  10882069     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We have determined the crystal structure of the enzyme-product complex of the hammerhead ribozyme by using a reinforced crystal lattice to trap the complex prior to dissociation and by employing X-ray holographic image reconstruction, a real-space electron density imaging and refinement procedure. Subsequent to catalysis, the cleavage site residue (C-17), together with its 2',3'-cyclic phosphate, adopts a conformation close to and approximately perpendicular to the Watson-Crick base-pairing faces of two highly conserved purines in the ribozyme's catalytic pocket (G-5 and A-6). We observe several interactions with functional groups on these residues that have been identified as critical for ribozyme activity by biochemical analyses but whose role has defied explanation in terms of previous structural analyses. These interactions may therefore be relevant to the hammerhead ribozyme reaction mechanism.
Authors:
J B Murray; H Szöke; A Szöke; W G Scott
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Molecular cell     Volume:  5     ISSN:  1097-2765     ISO Abbreviation:  Mol. Cell     Publication Date:  2000 Feb 
Date Detail:
Created Date:  2000-07-17     Completed Date:  2000-07-17     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  9802571     Medline TA:  Mol Cell     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  279-87     Citation Subset:  IM    
Affiliation:
The Center for the Molecular Biology of RNA and Department of Chemistry and Biochemistry, University of California, Santa Cruz 95064, USA.
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MeSH Terms
Descriptor/Qualifier:
Crystallography / methods
Holography / methods
Image Processing, Computer-Assisted / methods
Models, Molecular
Nucleic Acid Conformation
RNA, Catalytic / chemistry*
X-Rays
Grant Support
ID/Acronym/Agency:
1P41 RR12408-01A1/RR/NCRR NIH HHS; R01 AI43393-02/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/RNA, Catalytic

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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