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| - | '''Crystal Structure of a junctioned hairpin ribozyme incorporating 9atom linker and 2'-deoxy 2'-amino U at A-1'''
| + | ===Crystal Structure of a junctioned hairpin ribozyme incorporating 9atom linker and 2'-deoxy 2'-amino U at A-1=== |
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| - | ==Overview==
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| - | The hairpin ribozyme is a small catalytic RNA comprising two helix-loop-helix domains linked by a four-way helical junction (4WJ). In its most basic form, each domain can be formed independently and reconstituted without a 4WJ to yield an active enzyme. The production of such minimal junctionless hairpin ribozymes is achievable by chemical synthesis, which has allowed structures to be determined for numerous nucleotide variants. However, abasic and other destabilizing core modifications hinder crystallization. This investigation describes the use of a dangling 5'-U to form an intermolecular U.U mismatch, as well as the use of synthetic linkers to tether the loop A and B domains, including (i) a three-carbon propyl linker (C3L) and (ii) a nine-atom triethylene glycol linker (S9L). Both linker constructs demonstrated similar enzymatic activity, but S9L constructs yielded crystals that diffracted to 2.65 A resolution or better. In contrast, C3L variants diffracted to 3.35 A and exhibited a 15 A expansion of the c axis. Crystal packing of the C3L construct showed a paucity of 6(1) contacts, which comprise numerous backbone to 2'-OH hydrogen bonds in junctionless and S9L complexes. Significantly, the crystal packing in minimal structures mimics stabilizing features observed in the 4WJ hairpin ribozyme structure. The results demonstrate how knowledge-based design can be used to improve diffraction and overcome otherwise destabilizing defects.
| + | The line below this paragraph, {{ABSTRACT_PUBMED_17582172}}, adds the Publication Abstract to the page |
| | + | (as it appears on PubMed at http://www.pubmed.gov), where 17582172 is the PubMed ID number. |
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| | + | {{ABSTRACT_PUBMED_17582172}} |
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| | ==About this Structure== | | ==About this Structure== |
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| | [[Category: Ribose zipper]] | | [[Category: Ribose zipper]] |
| | [[Category: S-turn]] | | [[Category: S-turn]] |
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun May 4 09:45:45 2008'' | + | |
| | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Tue Jul 29 10:05:30 2008'' |
Revision as of 07:05, 29 July 2008
Template:STRUCTURE 2npy
Crystal Structure of a junctioned hairpin ribozyme incorporating 9atom linker and 2'-deoxy 2'-amino U at A-1
Template:ABSTRACT PUBMED 17582172
About this Structure
Full crystallographic information is available from OCA.
Reference
A posteriori design of crystal contacts to improve the X-ray diffraction properties of a small RNA enzyme., MacElrevey C, Spitale RC, Krucinska J, Wedekind JE, Acta Crystallogr D Biol Crystallogr. 2007 Jul;63(Pt 7):812-25. Epub 2007, Jun 15. PMID:17582172
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