Crystallization and preliminary X‐ray diffraction data of an LNA 7‐mer duplex derived from a ricin aptamer
Identifieur interne : 002902 ( Main/Exploration ); précédent : 002901; suivant : 002903Crystallization and preliminary X‐ray diffraction data of an LNA 7‐mer duplex derived from a ricin aptamer
Auteurs : Charlotte Förster ; Dominik Oberthuer ; Jiang Gao ; André Eichert ; Frederick G. Quast ; Christian Betzel ; Andreas Nitsche ; Volker A. Erdmann ; Jens P. FürsteSource :
- Acta Crystallographica Section F [ 1744-3091 ] ; 2009-09-01.
English descriptors
- Teeft :
- Acta, Acta cryst, Approximate dimensions, Aptamer, Aptamers, Asymmetric unit, Basic conditions, Biological activity, Cryst, Crystal growth, Crystal morphology, Crystal quality, Crystallization, Crystallization communications, Crystallization experiments, Crystallographic data, Different metal salts, Diffraction data, Duplex, Forster, Great extent, Hampton research, Helix, Linbro plates, Lithium sulfate, Lnas, Methylene, Methylene ribofuranose, Natural ribose, Nature biotechnol, Nucleic, Nucleic acids, Petersen, Preliminary diffraction data, Reservoir solution, Ribofuranose, Ricin, Ricin aptamer, Sodium cacodylate, Solvent content, Space group, Structure analysis, Tenascin aptamer, Terminal part, Unpublished results, Voss gerstein, Wengel, Znso4.
Abstract
Locked nucleic acids (LNAs) are modified nucleic acids which contain a modified sugar such as β‐d‐2′‐O,4′‐C methylene‐bridged ribofuranose or other sugar derivatives in LNA analogues. The β‐d‐2′‐O,4′‐C methylene ribofuranose LNAs in particular possess high stability and melting temperatures, which makes them of interest for stabilizing the structure of different nucleic acids. Aptamers, which are DNAs or RNAs targeted against specific ligands, are candidates for substitution with LNAs in order to increase their stability. A 7‐mer helix derived from the terminal part of an aptamer that was targeted against ricin was chosen. The ricin aptamer originally consisted of natural RNA building blocks and showed high affinity in ricin binding. For future stabilization of the aptamer, the terminal helix has been constructed as an `all‐locked' LNA and was successfully crystallized in order to investigate its structural properties. Optimization of crystal growth succeeded by the use of different metal salts as additives, such as CuCl2, MgCl2, MnCl2, CaCl2, CoCl2 and ZnSO4. Preliminary X‐ray diffraction data were collected and processed to 2.8 Å resolution. The LNA crystallized in space group P65, with unit‐cell parameters a = 50.11, b = 50.11, c = 40.72 Å. The crystals contained one LNA helix per asymmetric unit with a Matthews coefficient of 3.17 Å3 Da−1, which implies a solvent content of 70.15%.
Url:
DOI: 10.1107/S1744309109029145
Affiliations:
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Fürste</name></author></analytic><monogr></monogr><series><title level="j" type="main">Acta Crystallographica Section F</title><title level="j" type="alt">ACTA CRYSTALLOGRAPHICA F</title><idno type="ISSN">1744-3091</idno><idno type="eISSN">1744-3091</idno><imprint><biblScope unit="vol">65</biblScope><biblScope unit="issue">9</biblScope><biblScope unit="page" from="881">881</biblScope><biblScope unit="page" to="885">885</biblScope><publisher>International Union of Crystallography</publisher><pubPlace>5 Abbey Square, Chester, Cheshire CH1 2HU, England</pubPlace><date type="published" when="2009-09-01">2009-09-01</date></imprint><idno type="ISSN">1744-3091</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">1744-3091</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Acta</term><term>Acta cryst</term><term>Approximate dimensions</term><term>Aptamer</term><term>Aptamers</term><term>Asymmetric unit</term><term>Basic conditions</term><term>Biological activity</term><term>Cryst</term><term>Crystal growth</term><term>Crystal morphology</term><term>Crystal quality</term><term>Crystallization</term><term>Crystallization communications</term><term>Crystallization experiments</term><term>Crystallographic data</term><term>Different metal salts</term><term>Diffraction data</term><term>Duplex</term><term>Forster</term><term>Great extent</term><term>Hampton research</term><term>Helix</term><term>Linbro plates</term><term>Lithium sulfate</term><term>Lnas</term><term>Methylene</term><term>Methylene ribofuranose</term><term>Natural ribose</term><term>Nature biotechnol</term><term>Nucleic</term><term>Nucleic acids</term><term>Petersen</term><term>Preliminary diffraction data</term><term>Reservoir solution</term><term>Ribofuranose</term><term>Ricin</term><term>Ricin aptamer</term><term>Sodium cacodylate</term><term>Solvent content</term><term>Space group</term><term>Structure analysis</term><term>Tenascin aptamer</term><term>Terminal part</term><term>Unpublished results</term><term>Voss gerstein</term><term>Wengel</term><term>Znso4</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Locked nucleic acids (LNAs) are modified nucleic acids which contain a modified sugar such as β‐d‐2′‐O,4′‐C methylene‐bridged ribofuranose or other sugar derivatives in LNA analogues. The β‐d‐2′‐O,4′‐C methylene ribofuranose LNAs in particular possess high stability and melting temperatures, which makes them of interest for stabilizing the structure of different nucleic acids. Aptamers, which are DNAs or RNAs targeted against specific ligands, are candidates for substitution with LNAs in order to increase their stability. A 7‐mer helix derived from the terminal part of an aptamer that was targeted against ricin was chosen. The ricin aptamer originally consisted of natural RNA building blocks and showed high affinity in ricin binding. For future stabilization of the aptamer, the terminal helix has been constructed as an `all‐locked' LNA and was successfully crystallized in order to investigate its structural properties. Optimization of crystal growth succeeded by the use of different metal salts as additives, such as CuCl2, MgCl2, MnCl2, CaCl2, CoCl2 and ZnSO4. Preliminary X‐ray diffraction data were collected and processed to 2.8 Å resolution. The LNA crystallized in space group P65, with unit‐cell parameters a = 50.11, b = 50.11, c = 40.72 Å. The crystals contained one LNA helix per asymmetric unit with a Matthews coefficient of 3.17 Å3 Da−1, which implies a solvent content of 70.15%.</div></front></TEI><affiliations><list></list><tree><noCountry><name sortKey="Betzel, Christian" sort="Betzel, Christian" uniqKey="Betzel C" first="Christian" last="Betzel">Christian Betzel</name><name sortKey="Eichert, Andre" sort="Eichert, Andre" uniqKey="Eichert A" first="André" last="Eichert">André Eichert</name><name sortKey="Erdmann, Volker A" sort="Erdmann, Volker A" uniqKey="Erdmann V" first="Volker A." last="Erdmann">Volker A. Erdmann</name><name sortKey="Forster, Charlotte" sort="Forster, Charlotte" uniqKey="Forster C" first="Charlotte" last="Förster">Charlotte Förster</name><name sortKey="Furste, Jens P" sort="Furste, Jens P" uniqKey="Furste J" first="Jens P." last="Fürste">Jens P. Fürste</name><name sortKey="Gao, Jiang" sort="Gao, Jiang" uniqKey="Gao J" first="Jiang" last="Gao">Jiang Gao</name><name sortKey="Nitsche, Andreas" sort="Nitsche, Andreas" uniqKey="Nitsche A" first="Andreas" last="Nitsche">Andreas Nitsche</name><name sortKey="Oberthuer, Dominik" sort="Oberthuer, Dominik" uniqKey="Oberthuer D" first="Dominik" last="Oberthuer">Dominik Oberthuer</name><name sortKey="Quast, Frederick G" sort="Quast, Frederick G" uniqKey="Quast F" first="Frederick G." last="Quast">Frederick G. Quast</name></noCountry></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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