Mechanistic Studies Comparing the Incorporation of (+) and (−) Isomers of 3TCTP by HIV-1 Reverse Transcriptase†
Identifieur interne : 003842 ( Main/Exploration ); précédent : 003841; suivant : 003843Mechanistic Studies Comparing the Incorporation of (+) and (−) Isomers of 3TCTP by HIV-1 Reverse Transcriptase†
Auteurs : Joy Y. Feng [États-Unis] ; Karen S. Anderson [États-Unis]Source :
- Biochemistry [ 0006-2960 ] ; 1998.
Abstract
Among the nucleoside inhibitors used clinically as anti-HIV drugs which target HIV-1 reverse transcriptase (RT), (−)-2‘,3‘-dideoxy-3‘-thiacytidine [(−)SddC or 3TC] is the only analogue with the unnatural L(−) nucleoside configuration. 3TC has been shown to be more potent and less toxic than the D(+) isomer, (+)SddC, which has the natural nucleoside configuration. The mechanistic basis for the stereochemical selectivity and differential toxicity of the isomeric SddC compounds is not completely understood although a number of factors may clearly come into play including differences in uptake, metabolic activation, degradation, and transport. We used a pre-steady-state kinetic analysis to determine the maximum rate of incorporation, kpol, nucleotide-binding affinity, Kd, and efficiency of incorporation, kpol/Kd, for the (−) and (+) isomeric SddCTP compounds as well as the corresponding dideoxy and natural nucleoside triphosphates into a primer−template complex using HIV-1 reverse transcriptase. The affinity (Kd) of the dNTP was much tighter and the efficiency (kpol/Kd) of incorporation by enzyme into the primer−template complex was much higher for the DNA/RNA primer−template compared to DNA/DNA. The maximum rate of incorporation, kpol, followed the trend of dCTP > ddCTP > (+)SddCTP > (−)SddCTP while the Kd values determined for the DNA/RNA primer−template followed the order (−)SddCTP ≅ (+)SddCTP ≅ ddCTP > dCTP. The corresponding efficiency of incorporation followed the trend dCTP > ddCTP > (+)SddCTP > (−)SddCTP. These data suggest that perturbations on the ribose ring of cytidine analogues (C → S) decrease the rate and efficiency of incorporation but enhance the binding affinity. These results are discussed in the context of a computer modeled structure of the ternary complexes of RT, DNA/RNA primer−template, and SddCTP analogues as well as implications for structure−activity relationships and further drug design. This information provides a mechanistic basis for understanding the inhibition of HIV-1 reverse transcriptase by 3TC.
Url:
DOI: 10.1021/bi982340r
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: 002192
- to stream Istex, to step Curation: 002192
- to stream Istex, to step Checkpoint: 001095
- to stream Main, to step Merge: 003887
- to stream Main, to step Curation: 003842
Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title>Mechanistic Studies Comparing the Incorporation of (+) and (−) Isomers of 3TCTP by HIV-1 Reverse Transcriptase†</title><author><name sortKey="Feng, Joy Y" sort="Feng, Joy Y" uniqKey="Feng J" first="Joy Y." last="Feng">Joy Y. Feng</name></author><author><name sortKey="Anderson, Karen S" sort="Anderson, Karen S" uniqKey="Anderson K" first="Karen S." last="Anderson">Karen S. Anderson</name></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:414ADAB1978E8508E47426744DE36DE2F52FC93E</idno><date when="1999" year="1999">1999</date><idno type="doi">10.1021/bi982340r</idno><idno type="url">https://api.istex.fr/ark:/67375/TPS-MLHF9NSC-J/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">002192</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">002192</idno><idno type="wicri:Area/Istex/Curation">002192</idno><idno type="wicri:Area/Istex/Checkpoint">001095</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">001095</idno><idno type="wicri:doubleKey">0006-2960:1999:Feng J:mechanistic:studies:comparing</idno><idno type="wicri:Area/Main/Merge">003887</idno><idno type="wicri:Area/Main/Curation">003842</idno><idno type="wicri:Area/Main/Exploration">003842</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main">Mechanistic Studies Comparing the Incorporation of (+) and (−) Isomers of
3TCTP by HIV-1 Reverse Transcriptase<ref type="bib" target="#bi982340rAF2"><hi rend="superscript">†</hi></ref></title><author><name sortKey="Feng, Joy Y" sort="Feng, Joy Y" uniqKey="Feng J" first="Joy Y." last="Feng">Joy Y. Feng</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Connecticut</region></placeName><wicri:cityArea>Department of Pharmacology, Yale University School of Medicine, 333 Cedar Street, New Haven</wicri:cityArea></affiliation></author><author><name sortKey="Anderson, Karen S" sort="Anderson, Karen S" uniqKey="Anderson K" first="Karen S." last="Anderson">Karen S. Anderson</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Connecticut</region></placeName><wicri:cityArea>Department of Pharmacology, Yale University School of Medicine, 333 Cedar Street, New Haven</wicri:cityArea></affiliation><affiliation wicri:level="1"><country wicri:rule="url">États-Unis</country></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">Biochemistry</title><title level="j" type="abbrev">Biochemistry</title><idno type="ISSN">0006-2960</idno><idno type="eISSN">1520-4995</idno><imprint><publisher>American Chemical Society</publisher><date type="e-published">1998</date><date type="published">1999</date><biblScope unit="vol">38</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="55">55</biblScope><biblScope unit="page" to="63">63</biblScope></imprint><idno type="ISSN">0006-2960</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0006-2960</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract">Among the nucleoside inhibitors used clinically as anti-HIV drugs which target HIV-1 reverse transcriptase (RT), (−)-2‘,3‘-dideoxy-3‘-thiacytidine [(−)SddC or 3TC] is the only analogue with the unnatural L(−) nucleoside configuration. 3TC has been shown to be more potent and less toxic than the D(+) isomer, (+)SddC, which has the natural nucleoside configuration. The mechanistic basis for the stereochemical selectivity and differential toxicity of the isomeric SddC compounds is not completely understood although a number of factors may clearly come into play including differences in uptake, metabolic activation, degradation, and transport. We used a pre-steady-state kinetic analysis to determine the maximum rate of incorporation, kpol, nucleotide-binding affinity, Kd, and efficiency of incorporation, kpol/Kd, for the (−) and (+) isomeric SddCTP compounds as well as the corresponding dideoxy and natural nucleoside triphosphates into a primer−template complex using HIV-1 reverse transcriptase. The affinity (Kd) of the dNTP was much tighter and the efficiency (kpol/Kd) of incorporation by enzyme into the primer−template complex was much higher for the DNA/RNA primer−template compared to DNA/DNA. The maximum rate of incorporation, kpol, followed the trend of dCTP > ddCTP > (+)SddCTP > (−)SddCTP while the Kd values determined for the DNA/RNA primer−template followed the order (−)SddCTP ≅ (+)SddCTP ≅ ddCTP > dCTP. The corresponding efficiency of incorporation followed the trend dCTP > ddCTP > (+)SddCTP > (−)SddCTP. These data suggest that perturbations on the ribose ring of cytidine analogues (C → S) decrease the rate and efficiency of incorporation but enhance the binding affinity. These results are discussed in the context of a computer modeled structure of the ternary complexes of RT, DNA/RNA primer−template, and SddCTP analogues as well as implications for structure−activity relationships and further drug design. This information provides a mechanistic basis for understanding the inhibition of HIV-1 reverse transcriptase by 3TC.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Connecticut</li></region></list><tree><country name="États-Unis"><region name="Connecticut"><name sortKey="Feng, Joy Y" sort="Feng, Joy Y" uniqKey="Feng J" first="Joy Y." last="Feng">Joy Y. Feng</name></region><name sortKey="Anderson, Karen S" sort="Anderson, Karen S" uniqKey="Anderson K" first="Karen S." last="Anderson">Karen S. Anderson</name><name sortKey="Anderson, Karen S" sort="Anderson, Karen S" uniqKey="Anderson K" first="Karen S." last="Anderson">Karen S. Anderson</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 003842 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 003842 | 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:414ADAB1978E8508E47426744DE36DE2F52FC93E
|texte= Mechanistic Studies Comparing the Incorporation of (+) and (−) Isomers of 3TCTP by HIV-1 Reverse Transcriptase†
}}
| This area was generated with Dilib version V0.6.33. |  |