Sandbox Reserved 712: Difference between revisions
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<scene name='Sandbox_Reserved_712/L89v/1'>L89V</scene>. | <scene name='Sandbox_Reserved_712/L89v/1'>L89V</scene>. | ||
These mutations lead to a change in the susceptibility to the PI. In the case of 3ggu we observe a 32-fold susceptibility to [[darunavir]]. In comparison to | These mutations lead to a change in the susceptibility to the PI. In the case of 3ggu we observe a 32-fold susceptibility to [[darunavir]]. In comparison to [[amprenavir]], which is a structural related PI of darunavir, it only shows a 24-fold susceptibility. The key-mutations that are responsible for the darunavir resistance are V32I, I54L and I54M. Those were not found in PR<sub>DRV5</sub> which explains the smaller phenotypic changes in the susceptibility to darunavir. (Complete Table: [http://jvi.asm.org.scd-rproxy.u-strasbg.fr/content/83/17/8810/T4.expansion.html/ Genotypes and phenotype changes analyzed with recombinant virus assay]) Nevertheless, determining the inhibition constants by kinetic analysis using a chromogenic peptide substrate and the appropriate inhibitor, we can observe an increase of the K<sub>i</sub> value for all the samples in comparison to the wild-type virus. PR<sub>DRV5</sub> - which only has a specific activity of 5% of the wild-type value - also shows a smaller difference in k<sub>i</sub> value for darunavir in comparison to the other used samples. (Complete Table: [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2738195/table/t6/ K<sub>i</sub> values for the inhibitors of PR mutants]) <ref name="Molecular"> PMID:19535439 </ref> | ||
[[Image:Relative_vitality_values_for_recombinant_PRs_and_PRIs.jpg | thumb | 220px | left | Fig.1 Relative vitality values. <ref name="Molecular"/>]] | [[Image:Relative_vitality_values_for_recombinant_PRs_and_PRIs.jpg | thumb | 220px | left | Fig.1 Relative vitality values. <ref name="Molecular"/>]] | ||
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Despite the many mutations the k<sub>cat</sub> values were still between 30% and 50% of the wild-type value. In contrast the K<sub>m</sub> values of the mutants were (mostly) four- to eightfold higher than the wild-type PR. | Despite the many mutations the k<sub>cat</sub> values were still between 30% and 50% of the wild-type value. In contrast the K<sub>m</sub> values of the mutants were (mostly) four- to eightfold higher than the wild-type PR. | ||
(Complete Table: [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2738195/table/t5/ Enzyme characteristics of PR variants analyzed in this study]) <ref name="Molecular" /> | (Complete Table: [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2738195/table/t5/ Enzyme characteristics of PR variants analyzed in this study]) <ref name="Molecular" /> | ||
== '''Applications''' == | == '''Applications''' == | ||