Differential inhibitory activities and stabilisation of DNA aptamers against the SARS coronavirus helicase.
Identifieur interne : 001D92 ( Ncbi/Curation ); précédent : 001D91; suivant : 001D93Differential inhibitory activities and stabilisation of DNA aptamers against the SARS coronavirus helicase.
Auteurs : Ka To Shum [Hong Kong] ; Julian A. TannerSource :
- Chembiochem : a European journal of chemical biology [ 1439-7633 ] ; 2008.
Descripteurs français
- KwdFr :
- Aptamères nucléotidiques (), Aptamères nucléotidiques (pharmacologie), Cinétique, Concentration inhibitrice 50, G-quadruplexes, Helicase (antagonistes et inhibiteurs), Helicase (métabolisme), Protéines virales (antagonistes et inhibiteurs), Protéines virales (métabolisme), Séquence nucléotidique, Virus du SRAS (enzymologie).
- MESH :
- antagonistes et inhibiteurs : Helicase, Protéines virales.
- enzymologie : Virus du SRAS.
- métabolisme : Helicase, Protéines virales.
- pharmacologie : Aptamères nucléotidiques.
- Aptamères nucléotidiques, Cinétique, Concentration inhibitrice 50, G-quadruplexes, Séquence nucléotidique.
English descriptors
- KwdEn :
- Aptamers, Nucleotide (chemistry), Aptamers, Nucleotide (pharmacology), Base Sequence, DNA Helicases (antagonists & inhibitors), DNA Helicases (metabolism), G-Quadruplexes, Inhibitory Concentration 50, Kinetics, SARS Virus (enzymology), Viral Proteins (antagonists & inhibitors), Viral Proteins (metabolism).
- MESH :
- chemical , antagonists & inhibitors : DNA Helicases, Viral Proteins.
- chemical , chemistry : Aptamers, Nucleotide.
- chemical , metabolism : DNA Helicases, Viral Proteins.
- chemical , pharmacology : Aptamers, Nucleotide.
- enzymology : SARS Virus.
- Base Sequence, G-Quadruplexes, Inhibitory Concentration 50, Kinetics.
Abstract
The helicase from severe acute respiratory syndrome coronavirus (SARS-CoV) possesses NTPase, duplex RNA/DNA-unwinding and RNA-capping activities that are essential for viral replication and proliferation. Here, we have isolated DNA aptamers against the SARS-CoV helicase from a combinatorial DNA library. These aptamers show two distinct classes of secondary structure, G-quadruplex and non-G-quadruplex, as shown by circular dichroism and gel electrophoresis. All of the aptamers that were selected stimulated ATPase activity of the SARS-CoV helicase with low-nanomolar apparent K(m) values. Intriguingly, only the non-G-quadruplex aptamers showed specific inhibition of helicase activities, whereas the G-quadruplex aptamers did not inhibit helicase activities. The non-G-quadruplex aptamer with the strongest inhibitory potency was modified at the 3'-end with biotin or inverted thymidine, and the modification increased its stability in serum, particularly for the inverted thymidine modification. Structural diversity in selection coupled to post-selection stabilisation has provided new insights into the aptamers that were selected for a helicase target. These aptamers are being further developed to inhibit SARS-CoV replication.
DOI: 10.1002/cbic.200800491
PubMed: 19031435
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Tanner</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2008">2008</date><idno type="RBID">pubmed:19031435</idno><idno type="pmid">19031435</idno><idno type="doi">10.1002/cbic.200800491</idno><idno type="wicri:Area/PubMed/Corpus">001A28</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001A28</idno><idno type="wicri:Area/PubMed/Curation">001A28</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">001A28</idno><idno type="wicri:Area/PubMed/Checkpoint">001B46</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">001B46</idno><idno type="wicri:Area/Ncbi/Merge">001D92</idno><idno type="wicri:Area/Ncbi/Curation">001D92</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Differential inhibitory activities and stabilisation of DNA aptamers against the SARS coronavirus helicase.</title><author><name sortKey="Shum, Ka To" sort="Shum, Ka To" uniqKey="Shum K" first="Ka To" last="Shum">Ka To Shum</name><affiliation wicri:level="1"><nlm:affiliation>Department of Biochemistry, University of Hong Kong, Pokfulam, Hong Kong.</nlm:affiliation><country xml:lang="fr">Hong Kong</country><wicri:regionArea>Department of Biochemistry, University of Hong Kong, Pokfulam</wicri:regionArea><wicri:noRegion>Pokfulam</wicri:noRegion></affiliation></author><author><name sortKey="Tanner, Julian A" sort="Tanner, Julian A" uniqKey="Tanner J" first="Julian A" last="Tanner">Julian A. Tanner</name></author></analytic><series><title level="j">Chembiochem : a European journal of chemical biology</title><idno type="eISSN">1439-7633</idno><imprint><date when="2008" type="published">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Aptamers, Nucleotide (chemistry)</term><term>Aptamers, Nucleotide (pharmacology)</term><term>Base Sequence</term><term>DNA Helicases (antagonists & inhibitors)</term><term>DNA Helicases (metabolism)</term><term>G-Quadruplexes</term><term>Inhibitory Concentration 50</term><term>Kinetics</term><term>SARS Virus (enzymology)</term><term>Viral Proteins (antagonists & inhibitors)</term><term>Viral Proteins (metabolism)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Aptamères nucléotidiques ()</term><term>Aptamères nucléotidiques (pharmacologie)</term><term>Cinétique</term><term>Concentration inhibitrice 50</term><term>G-quadruplexes</term><term>Helicase (antagonistes et inhibiteurs)</term><term>Helicase (métabolisme)</term><term>Protéines virales (antagonistes et inhibiteurs)</term><term>Protéines virales (métabolisme)</term><term>Séquence nucléotidique</term><term>Virus du SRAS (enzymologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="antagonists & inhibitors" xml:lang="en"><term>DNA Helicases</term><term>Viral Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Aptamers, Nucleotide</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>DNA Helicases</term><term>Viral Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Aptamers, Nucleotide</term></keywords><keywords scheme="MESH" qualifier="antagonistes et inhibiteurs" xml:lang="fr"><term>Helicase</term><term>Protéines virales</term></keywords><keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Helicase</term><term>Protéines virales</term></keywords><keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>Aptamères nucléotidiques</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Base Sequence</term><term>G-Quadruplexes</term><term>Inhibitory Concentration 50</term><term>Kinetics</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Aptamères nucléotidiques</term><term>Cinétique</term><term>Concentration inhibitrice 50</term><term>G-quadruplexes</term><term>Séquence nucléotidique</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The helicase from severe acute respiratory syndrome coronavirus (SARS-CoV) possesses NTPase, duplex RNA/DNA-unwinding and RNA-capping activities that are essential for viral replication and proliferation. Here, we have isolated DNA aptamers against the SARS-CoV helicase from a combinatorial DNA library. These aptamers show two distinct classes of secondary structure, G-quadruplex and non-G-quadruplex, as shown by circular dichroism and gel electrophoresis. All of the aptamers that were selected stimulated ATPase activity of the SARS-CoV helicase with low-nanomolar apparent K(m) values. Intriguingly, only the non-G-quadruplex aptamers showed specific inhibition of helicase activities, whereas the G-quadruplex aptamers did not inhibit helicase activities. The non-G-quadruplex aptamer with the strongest inhibitory potency was modified at the 3'-end with biotin or inverted thymidine, and the modification increased its stability in serum, particularly for the inverted thymidine modification. Structural diversity in selection coupled to post-selection stabilisation has provided new insights into the aptamers that were selected for a helicase target. These aptamers are being further developed to inhibit SARS-CoV replication.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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