Rapid determination of the affinity of 28- and 14-mer phosphorothioate oligonucleotides for HIV-1 reverse transcriptase by fluorescence spectroscopy
Identifieur interne : 004549 ( Main/Exploration ); précédent : 004548; suivant : 004550Rapid determination of the affinity of 28- and 14-mer phosphorothioate oligonucleotides for HIV-1 reverse transcriptase by fluorescence spectroscopy
Auteurs : Georges Maury [France] ; Gilles Divita [Allemagne] ; François Morvan [France] ; Jean-Louis Imbach [France] ; Roger S. Goody [Allemagne]Source :
- BBA - Gene Structure and Expression [ 0167-4781 ] ; 1993.
English descriptors
- Teeft :
- Affinity, Analog, Corresponding dissociation constants, Corresponding hybrids, Different methods, Displacement experiments, Dissociation constants, Duplex, Excitation, First step, Fluorescence, Fluorescence changes, Fluorescence emission, Fluorescence emission intensity, Fluorescence intensity, Fluorescence measurements, Fluorescence spectrometry, Fluorescence spectroscopy, Fluorescent ligand, Fluorescent template, Hybrid, Independent methods, Inhibition constants, Intrinsic fluorescence, Intrinsic fluorescence changes, Intrinsic fluorescence intensity, Ligand, Ligand affinities, Little influence, Mcmillan press, Normal oligomers, Normal oligonucleotides, Normal series, Nucleic acids, Oligomer, Oligomers, Oligonucleotide, Oligonucleotides, Phosphorothioate, Phosphorothioate cytidinylate oligonucleotides, Phosphorothioate oligocytidinylates, Phosphorothioate oligodeoxycytidinylates, Phosphorothioate oligomers, Phosphorothioate oligonucleotide, Phosphorothioate oligonucleotides, Present work, Primer, Primer analog, Primer hybrids, Quadratic equation, Rain incubation, Relative changes, Same conditions, Saturating concentration, Saturating concentrations, Several types, Steady state, Steady state kinetics, Structural biology, Template, Template primer, Tetrahedron lett, Titration, Transcriptase.
Abstract
Abstract: Intrinsic fluorescence of human immunodeficiency virus type 1 reverse transcriptase (E.C. 2.7.7.49) and displacement experiments of a fluorescent template · primer probe were used to study the interaction of the enzyme with several types of 28-and 14-mer normal or phosphorothioate oligodeoxycytidinylates and their duplexes with poly(rI). The two methods gave convergent results and allowed in each case fast determinations of ligand affinities for the enzyme. The dissociation constants (Kd) obtained from intrinsic fluorescence changes were slightly lower than those determined from the less direct competitive displacement experiments. In all cases, the enzyme displayed better recognition of the hybrid than of the unannealed oligonucleotide. The Kd values of phosphorothioate oligomers and their hybrids were lower than those of the corresponding normal oligomers and hybrids, but the difference was not as significant as in the case of the Ki constants for (dC)28 and S(dC)28 (Majumdar et al. (1989) Biochemistry 28, 1340). The affinities of the annealed phosphorothioate oligodeoxycytidinylates for the enzyme were found to be larger than for any other compounds in this series (Kd of poly(rI) · S(dC)28: 0.28 nM at 25°C). Changing the β stereochemistry of the oligomer bases to α did not alter the affinity of the oligodeoxycytidinylate and its hybrids for the enzyme.
Url:
DOI: 10.1016/0167-4781(93)90030-H
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: 000144
- to stream Istex, to step Curation: 000144
- to stream Istex, to step Checkpoint: 001B60
- to stream Main, to step Merge: 004615
- to stream Main, to step Curation: 004549
Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title>Rapid determination of the affinity of 28- and 14-mer phosphorothioate oligonucleotides for HIV-1 reverse transcriptase by fluorescence spectroscopy</title><author><name sortKey="Maury, Georges" sort="Maury, Georges" uniqKey="Maury G" first="Georges" last="Maury">Georges Maury</name></author><author><name sortKey="Divita, Gilles" sort="Divita, Gilles" uniqKey="Divita G" first="Gilles" last="Divita">Gilles Divita</name></author><author><name sortKey="Morvan, Francois" sort="Morvan, Francois" uniqKey="Morvan F" first="François" last="Morvan">François Morvan</name></author><author><name sortKey="Imbach, Jean Louis" sort="Imbach, Jean Louis" uniqKey="Imbach J" first="Jean-Louis" last="Imbach">Jean-Louis Imbach</name></author><author><name sortKey="Goody, Roger S" sort="Goody, Roger S" uniqKey="Goody R" first="Roger S." last="Goody">Roger S. Goody</name></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:D4BB46D332EB7C060AFB2594A222668376B93D80</idno><date when="1993" year="1993">1993</date><idno type="doi">10.1016/0167-4781(93)90030-H</idno><idno type="url">https://api.istex.fr/ark:/67375/6H6-6702S3R2-B/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">000144</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">000144</idno><idno type="wicri:Area/Istex/Curation">000144</idno><idno type="wicri:Area/Istex/Checkpoint">001B60</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">001B60</idno><idno type="wicri:doubleKey">0167-4781:1993:Maury G:rapid:determination:of</idno><idno type="wicri:Area/Main/Merge">004615</idno><idno type="wicri:Area/Main/Curation">004549</idno><idno type="wicri:Area/Main/Exploration">004549</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a">Rapid determination of the affinity of 28- and 14-mer phosphorothioate oligonucleotides for HIV-1 reverse transcriptase by fluorescence spectroscopy</title><author><name sortKey="Maury, Georges" sort="Maury, Georges" uniqKey="Maury G" first="Georges" last="Maury">Georges Maury</name><affiliation wicri:level="1"><country xml:lang="fr">France</country><wicri:regionArea>Université de Montpellier II, URA 488 du C.N.R.S.</wicri:regionArea><wicri:noRegion>URA 488 du C.N.R.S.</wicri:noRegion><wicri:noRegion>URA 488 du C.N.R.S.</wicri:noRegion></affiliation></author><author><name sortKey="Divita, Gilles" sort="Divita, Gilles" uniqKey="Divita G" first="Gilles" last="Divita">Gilles Divita</name><affiliation wicri:level="1"><country xml:lang="fr">Allemagne</country><wicri:regionArea>Abteilung Biophysik, Max-Planck-Institut für medizinische Forschung</wicri:regionArea><wicri:noRegion>Max-Planck-Institut für medizinische Forschung</wicri:noRegion><wicri:noRegion>Max-Planck-Institut für medizinische Forschung</wicri:noRegion></affiliation></author><author><name sortKey="Morvan, Francois" sort="Morvan, Francois" uniqKey="Morvan F" first="François" last="Morvan">François Morvan</name><affiliation wicri:level="1"><country xml:lang="fr">France</country><wicri:regionArea>Université de Montpellier II, URA 488 du C.N.R.S.</wicri:regionArea><wicri:noRegion>URA 488 du C.N.R.S.</wicri:noRegion><wicri:noRegion>URA 488 du C.N.R.S.</wicri:noRegion></affiliation></author><author><name sortKey="Imbach, Jean Louis" sort="Imbach, Jean Louis" uniqKey="Imbach J" first="Jean-Louis" last="Imbach">Jean-Louis Imbach</name><affiliation wicri:level="1"><country xml:lang="fr">France</country><wicri:regionArea>Université de Montpellier II, URA 488 du C.N.R.S.</wicri:regionArea><wicri:noRegion>URA 488 du C.N.R.S.</wicri:noRegion><wicri:noRegion>URA 488 du C.N.R.S.</wicri:noRegion></affiliation></author><author><name sortKey="Goody, Roger S" sort="Goody, Roger S" uniqKey="Goody R" first="Roger S." last="Goody">Roger S. Goody</name><affiliation wicri:level="1"><country xml:lang="fr">Allemagne</country><wicri:regionArea>Abteilung Biophysik, Max-Planck-Institut für medizinische Forschung</wicri:regionArea><wicri:noRegion>Max-Planck-Institut für medizinische Forschung</wicri:noRegion><wicri:noRegion>Max-Planck-Institut für medizinische Forschung</wicri:noRegion></affiliation></author></analytic><monogr></monogr><series><title level="j">BBA - Gene Structure and Expression</title><title level="j" type="abbrev">BBAEXP</title><idno type="ISSN">0167-4781</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1993">1993</date><biblScope unit="volume">1216</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="1">1</biblScope><biblScope unit="page" to="8">8</biblScope></imprint><idno type="ISSN">0167-4781</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0167-4781</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Affinity</term><term>Analog</term><term>Corresponding dissociation constants</term><term>Corresponding hybrids</term><term>Different methods</term><term>Displacement experiments</term><term>Dissociation constants</term><term>Duplex</term><term>Excitation</term><term>First step</term><term>Fluorescence</term><term>Fluorescence changes</term><term>Fluorescence emission</term><term>Fluorescence emission intensity</term><term>Fluorescence intensity</term><term>Fluorescence measurements</term><term>Fluorescence spectrometry</term><term>Fluorescence spectroscopy</term><term>Fluorescent ligand</term><term>Fluorescent template</term><term>Hybrid</term><term>Independent methods</term><term>Inhibition constants</term><term>Intrinsic fluorescence</term><term>Intrinsic fluorescence changes</term><term>Intrinsic fluorescence intensity</term><term>Ligand</term><term>Ligand affinities</term><term>Little influence</term><term>Mcmillan press</term><term>Normal oligomers</term><term>Normal oligonucleotides</term><term>Normal series</term><term>Nucleic acids</term><term>Oligomer</term><term>Oligomers</term><term>Oligonucleotide</term><term>Oligonucleotides</term><term>Phosphorothioate</term><term>Phosphorothioate cytidinylate oligonucleotides</term><term>Phosphorothioate oligocytidinylates</term><term>Phosphorothioate oligodeoxycytidinylates</term><term>Phosphorothioate oligomers</term><term>Phosphorothioate oligonucleotide</term><term>Phosphorothioate oligonucleotides</term><term>Present work</term><term>Primer</term><term>Primer analog</term><term>Primer hybrids</term><term>Quadratic equation</term><term>Rain incubation</term><term>Relative changes</term><term>Same conditions</term><term>Saturating concentration</term><term>Saturating concentrations</term><term>Several types</term><term>Steady state</term><term>Steady state kinetics</term><term>Structural biology</term><term>Template</term><term>Template primer</term><term>Tetrahedron lett</term><term>Titration</term><term>Transcriptase</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Intrinsic fluorescence of human immunodeficiency virus type 1 reverse transcriptase (E.C. 2.7.7.49) and displacement experiments of a fluorescent template · primer probe were used to study the interaction of the enzyme with several types of 28-and 14-mer normal or phosphorothioate oligodeoxycytidinylates and their duplexes with poly(rI). The two methods gave convergent results and allowed in each case fast determinations of ligand affinities for the enzyme. The dissociation constants (Kd) obtained from intrinsic fluorescence changes were slightly lower than those determined from the less direct competitive displacement experiments. In all cases, the enzyme displayed better recognition of the hybrid than of the unannealed oligonucleotide. The Kd values of phosphorothioate oligomers and their hybrids were lower than those of the corresponding normal oligomers and hybrids, but the difference was not as significant as in the case of the Ki constants for (dC)28 and S(dC)28 (Majumdar et al. (1989) Biochemistry 28, 1340). The affinities of the annealed phosphorothioate oligodeoxycytidinylates for the enzyme were found to be larger than for any other compounds in this series (Kd of poly(rI) · S(dC)28: 0.28 nM at 25°C). Changing the β stereochemistry of the oligomer bases to α did not alter the affinity of the oligodeoxycytidinylate and its hybrids for the enzyme.</div></front></TEI><affiliations><list><country><li>Allemagne</li><li>France</li></country></list><tree><country name="France"><noRegion><name sortKey="Maury, Georges" sort="Maury, Georges" uniqKey="Maury G" first="Georges" last="Maury">Georges Maury</name></noRegion><name sortKey="Imbach, Jean Louis" sort="Imbach, Jean Louis" uniqKey="Imbach J" first="Jean-Louis" last="Imbach">Jean-Louis Imbach</name><name sortKey="Morvan, Francois" sort="Morvan, Francois" uniqKey="Morvan F" first="François" last="Morvan">François Morvan</name></country><country name="Allemagne"><noRegion><name sortKey="Divita, Gilles" sort="Divita, Gilles" uniqKey="Divita G" first="Gilles" last="Divita">Gilles Divita</name></noRegion><name sortKey="Goody, Roger S" sort="Goody, Roger S" uniqKey="Goody R" first="Roger S." last="Goody">Roger S. Goody</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 004549 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 004549 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= MersV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= ISTEX:D4BB46D332EB7C060AFB2594A222668376B93D80
|texte= Rapid determination of the affinity of 28- and 14-mer phosphorothioate oligonucleotides for HIV-1 reverse transcriptase by fluorescence spectroscopy
}}
| This area was generated with Dilib version V0.6.33. |  |