Capturing hammerhead ribozyme structures in action by modulating general base catalysis.
We have obtained precatalytic (enzyme-substrate complex) and postcatalytic (enzyme-product complex) crystal structures of an active full-length hammerhead RNA that cleaves in the crystal. Using the natural satellite tobacco ringspot virus hammerhead RNA sequence, the self-cleavage reaction was modul...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2008-09-01
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| Series: | PLoS Biology |
| Online Access: | http://europepmc.org/articles/PMC2553840?pdf=render |
| _version_ | 1818622926814969856 |
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| author | Young-In Chi Monika Martick Monica Lares Rosalind Kim William G Scott Sung-Hou Kim |
| author_facet | Young-In Chi Monika Martick Monica Lares Rosalind Kim William G Scott Sung-Hou Kim |
| author_sort | Young-In Chi |
| collection | DOAJ |
| description | We have obtained precatalytic (enzyme-substrate complex) and postcatalytic (enzyme-product complex) crystal structures of an active full-length hammerhead RNA that cleaves in the crystal. Using the natural satellite tobacco ringspot virus hammerhead RNA sequence, the self-cleavage reaction was modulated by substituting the general base of the ribozyme, G12, with A12, a purine variant with a much lower pKa that does not significantly perturb the ribozyme's atomic structure. The active, but slowly cleaving, ribozyme thus permitted isolation of enzyme-substrate and enzyme-product complexes without modifying the nucleophile or leaving group of the cleavage reaction, nor any other aspect of the substrate. The predissociation enzyme-product complex structure reveals RNA and metal ion interactions potentially relevant to transition-state stabilization that are absent in precatalytic structures. |
| first_indexed | 2024-12-16T18:32:56Z |
| format | Article |
| id | doaj.art-2c7eee625ab74664839655b8acde31a2 |
| institution | Directory Open Access Journal |
| issn | 1544-9173 1545-7885 |
| language | English |
| last_indexed | 2024-12-16T18:32:56Z |
| publishDate | 2008-09-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Biology |
| spelling | doaj.art-2c7eee625ab74664839655b8acde31a22022-12-21T22:21:14ZengPublic Library of Science (PLoS)PLoS Biology1544-91731545-78852008-09-0169e23410.1371/journal.pbio.0060234Capturing hammerhead ribozyme structures in action by modulating general base catalysis.Young-In ChiMonika MartickMonica LaresRosalind KimWilliam G ScottSung-Hou KimWe have obtained precatalytic (enzyme-substrate complex) and postcatalytic (enzyme-product complex) crystal structures of an active full-length hammerhead RNA that cleaves in the crystal. Using the natural satellite tobacco ringspot virus hammerhead RNA sequence, the self-cleavage reaction was modulated by substituting the general base of the ribozyme, G12, with A12, a purine variant with a much lower pKa that does not significantly perturb the ribozyme's atomic structure. The active, but slowly cleaving, ribozyme thus permitted isolation of enzyme-substrate and enzyme-product complexes without modifying the nucleophile or leaving group of the cleavage reaction, nor any other aspect of the substrate. The predissociation enzyme-product complex structure reveals RNA and metal ion interactions potentially relevant to transition-state stabilization that are absent in precatalytic structures.http://europepmc.org/articles/PMC2553840?pdf=render |
| spellingShingle | Young-In Chi Monika Martick Monica Lares Rosalind Kim William G Scott Sung-Hou Kim Capturing hammerhead ribozyme structures in action by modulating general base catalysis. PLoS Biology |
| title | Capturing hammerhead ribozyme structures in action by modulating general base catalysis. |
| title_full | Capturing hammerhead ribozyme structures in action by modulating general base catalysis. |
| title_fullStr | Capturing hammerhead ribozyme structures in action by modulating general base catalysis. |
| title_full_unstemmed | Capturing hammerhead ribozyme structures in action by modulating general base catalysis. |
| title_short | Capturing hammerhead ribozyme structures in action by modulating general base catalysis. |
| title_sort | capturing hammerhead ribozyme structures in action by modulating general base catalysis |
| url | http://europepmc.org/articles/PMC2553840?pdf=render |
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