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Base-specific spin labeling of RNA for structure determination
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dc.contributor.author | Piton, N. | |
dc.contributor.author | Mu, Y. | |
dc.contributor.author | Stock, G. | |
dc.contributor.author | Prisner, TF. | |
dc.contributor.author | Schiemann, Olav | |
dc.contributor.author | Engels, J.W. | |
dc.date.accessioned | 2013-12-05T13:01:03Z | |
dc.date.available | 2013-12-05T13:01:03Z | |
dc.date.issued | 2007-05 | |
dc.identifier.citation | Piton , N , Mu , Y , Stock , G , Prisner , TF , Schiemann , O & Engels , J W 2007 , ' Base-specific spin labeling of RNA for structure determination ' , Nucleic Acids Research , vol. 35 , no. 9 , pp. 3128-3143 . https://doi.org/10.1093/nar/gkm169 | en |
dc.identifier.issn | 0305-1048 | |
dc.identifier.other | PURE: 392230 | |
dc.identifier.other | PURE UUID: 39254a48-0200-4373-8c3c-3adb2954ead9 | |
dc.identifier.other | WOS: 000247350800030 | |
dc.identifier.other | Scopus: 34250688400 | |
dc.identifier.uri | http://hdl.handle.net/10023/4274 | |
dc.description.abstract | To facilitate the measurement of intramolecular distances in solvated RNA systems, a combination of spin- labeling, electron paramagnetic resonance ( EPR), and molecular dynamics ( MD) simulation is presented. The fairly rigid spin label 2,2,5,5- tetramethylpyrrolin- 1- yloxyl- 3- acetylene ( TPA) was base and site specifically introduced into RNA through a Sonogashira palladium catalyzed crosscoupling on column. For this purpose 5- iodouridine, 5- iodo- cytidine and 2- iodo- adenosine phosphoramidites were synthesized and incorporated into RNA- sequences. Application of the recently developed ACE (R) chemistry presented the main advantage to limit the reduction of the nitroxide to an amine during the oligonucleotide automated synthesis and thus to increase substantially the reliability of the synthesis and the yield of labeled oligonucleotides. 4- Pulse Electron Double Resonance ( PELDOR) was then successfully used to measure the intramolecular spin - spin distances in six doubly labeled RNA- duplexes. Comparison of these results with our previous work on DNA showed that A- and B- Form can be differentiated. Using an all- atom force field with explicit solvent, MD simulations gave results in good agreement with the measured distances and indicated that the RNA A- Form was conserved despite a local destabilization effect of the nitroxide label. The applicability of the method to more complex biological systems is discussed. | |
dc.format.extent | 16 | |
dc.language.iso | eng | |
dc.relation.ispartof | Nucleic Acids Research | en |
dc.rights | © 2007 The Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. | en |
dc.subject | PAIR DISTANCE DISTRIBUTIONS | en |
dc.subject | EPR SPECTROSCOPY | en |
dc.subject | NUCLEIC-ACIDS | en |
dc.subject | TIKHONOV REGULARIZATION | en |
dc.subject | PARAMAGNETIC-RESONANCE | en |
dc.subject | HAMMERHEAD RIBOZYME | en |
dc.subject | PROTECTING GROUPS | en |
dc.subject | PULSED ESR | en |
dc.title | Base-specific spin labeling of RNA for structure determination | en |
dc.type | Journal article | en |
dc.description.version | Publisher PDF | en |
dc.contributor.institution | University of St Andrews. School of Biology | en |
dc.contributor.institution | University of St Andrews. Biomedical Sciences Research Complex | en |
dc.identifier.doi | https://doi.org/10.1093/nar/gkm169 | |
dc.description.status | Peer reviewed | en |
dc.identifier.url | http://www.scopus.com/inward/record.url?scp=34250688400&partnerID=8YFLogxK | en |
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