Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) in inclusion bodies | Human immunodeficiency virus 1 |
Crystallization (Comment) | Organism |
---|---|
purified recombinant wild-type enzyme in complex with inhibitors ritonavir or indinavir, hanging drop vapour diffusion method, 3 mg/ml protein in 50 mM sodium acetate, pH 4.7, is mixed in a ratio of 10:1 with inhibitor ritonavir or indinavir solubilized in 100% DMSO, protein-inhibitor slution is mixed with an equal volume of reservoir solution, containing 1.5 M ammonium sulfate and 20 mM sodium acetate at pH 6.0 giving hanging drops of 0.004 ml, room temperature, one week, X-ray diffraction and analysis at 1.9-2.5 A resolution | Human immunodeficiency virus 1 |
Protein Variants | Comment | Organism |
---|---|---|
A71V | site-directed mutagenesis of wild-type V6 enzyme, non-active-site mutation, inhibitor binding structure analysis | Human immunodeficiency virus 1 |
I54V | site-directed mutagenesis of wild-type V6 enzyme, non-active-site mutation, the mutation leads to resistance against ritonavir, inhibitor binding structure analysis | Human immunodeficiency virus 1 |
I54V/I84V | site-directed mutagenesis of wild-type V6 enzyme, non-active-site mutation, the mutation leads to resistance against ritonavir, inhibitor binding structure analysis | Human immunodeficiency virus 1 |
I84V | site-directed mutagenesis of wild-type V6 enzyme, active-site mutation, the mutation leads to resistance against ritonavir, the mutant shows 10fold reduced catalytic effciency compared to the wild-type enzyme, inhibitor binding structure analysis | Human immunodeficiency virus 1 |
K20R | site-directed mutagenesis of wild-type V6 enzyme, non-active-site mutation, inhibitor binding structure analysis | Human immunodeficiency virus 1 |
L33F | site-directed mutagenesis of wild-type V6 enzyme, non-active-site mutation, inhibitor binding structure analysis | Human immunodeficiency virus 1 |
L63P | site-directed mutagenesis of wild-type V6 enzyme, non-active-site mutation, inhibitor binding structure analysis | Human immunodeficiency virus 1 |
L90M | site-directed mutagenesis of wild-type V6 enzyme, non-active-site mutation, inhibitor binding structure analysis | Human immunodeficiency virus 1 |
M36I | site-directed mutagenesis of wild-type V6 enzyme, non-active-site mutation, inhibitor binding structure analysis | Human immunodeficiency virus 1 |
M46I | site-directed mutagenesis of wild-type V6 enzyme, non-active-site mutation, the mutation leads to resistance against ritonavir, inhibitor binding structure analysis | Human immunodeficiency virus 1 |
M46I/I54V/I84V | site-directed mutagenesis of wild-type V6 enzyme, non-active-site mutation, the mutation leads to resistance against ritonavir, inhibitor binding structure analysis | Human immunodeficiency virus 1 |
M46I/I84V | site-directed mutagenesis of wild-type V6 enzyme, non-active-site mutation, the mutation leads to resistance against ritonavir, inhibitor binding structure analysis | Human immunodeficiency virus 1 |
V32I | site-directed mutagenesis of wild-type V6 enzyme, the mutation leads to resistance against ritonavir, inhibitor binding structure analysis | Human immunodeficiency virus 1 |
V82A | site-directed mutagenesis of wild-type V6 enzyme, the mutation leads to resistance against ritonavir, inhibitor binding structure analysis | Human immunodeficiency virus 1 |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
AG1776 | - |
Human immunodeficiency virus 1 | |
indinavir | - |
Human immunodeficiency virus 1 | |
additional information | active site inhibitor binding structure analysis, overview | Human immunodeficiency virus 1 | |
nelfinavir | - |
Human immunodeficiency virus 1 | |
ritonavir | - |
Human immunodeficiency virus 1 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics, LAI wild-type enzyme | Human immunodeficiency virus 1 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Human immunodeficiency virus 1 | - |
i.e. HIV-1 | - |
Purification (Comment) | Organism |
---|---|
refolded recombinant wild-type and mutant enzymes by ammonium sulfate precipitation and gel filtration | Human immunodeficiency virus 1 |
Renatured (Comment) | Organism |
---|---|
solubilization by 8 M urea and renaturation by dialysis against 0.05 M sodium phosphate, containing 5 mM EDTA, 0.3 M NaCl, and 1 mM DTT | Human immunodeficiency virus 1 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
Lys-Ala-Arg-Val-Leu-4-nitro-Phe-Glu-Ala-Nle-Gly + H2O | the peptide substrate mimicks the CA-p2 cleavage site | Human immunodeficiency virus 1 | Lys-Ala-Arg-Val-Leu + 4-nitro-Phe-Glu-Ala-Nle-Gly | - |
? |
Synonyms | Comment | Organism |
---|---|---|
HIV-1 PR | - |
Human immunodeficiency virus 1 |
HIV-1 protease | - |
Human immunodeficiency virus 1 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Human immunodeficiency virus 1 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
4.7 | - |
assay at | Human immunodeficiency virus 1 |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.000000021 | - |
AG1776 | pH 4.7, 37°C, recombinant LAI wild-type enzyme | Human immunodeficiency virus 1 | |
0.00000004 | - |
AG1776 | pH 4.7, 37°C, recombinant V6 wild-type enzyme | Human immunodeficiency virus 1 | |
0.00000019 | - |
AG1776 | pH 4.7, 37°C, recombinant V6 mutant I54V | Human immunodeficiency virus 1 | |
0.0000005 | - |
AG1776 | pH 4.7, 37°C, recombinant V6 mutant M46I | Human immunodeficiency virus 1 | |
0.00000055 | - |
AG1776 | pH 4.7, 37°C, recombinant V6 mutant I84V | Human immunodeficiency virus 1 | |
0.0000007 | - |
ritonavir | pH 4.7, 37°C, recombinant LAI wild-type enzyme | Human immunodeficiency virus 1 | |
0.0000012 | - |
nelfinavir | pH 4.7, 37°C, recombinant LAI wild-type enzyme | Human immunodeficiency virus 1 | |
0.0000022 | - |
AG1776 | pH 4.7, 37°C, recombinant V6 mutant M46I/I84V | Human immunodeficiency virus 1 | |
0.000003 | - |
ritonavir | pH 4.7, 37°C, recombinant V6 wild-type enzyme | Human immunodeficiency virus 1 | |
0.0000031 | - |
indinavir | pH 4.7, 37°C, recombinant LAI wild-type enzyme | Human immunodeficiency virus 1 | |
0.0000056 | - |
AG1776 | pH 4.7, 37°C, recombinant V6 mutant M46I/I54V/I84V | Human immunodeficiency virus 1 | |
0.0000065 | - |
ritonavir | pH 4.7, 37°C, recombinant V6 mutant M46I | Human immunodeficiency virus 1 | |
0.0000094 | - |
AG1776 | pH 4.7, 37°C, recombinant V6 mutant I54V/I84V | Human immunodeficiency virus 1 | |
0.0000109 | - |
ritonavir | pH 4.7, 37°C, recombinant V6 mutant I54V | Human immunodeficiency virus 1 | |
0.0000165 | - |
ritonavir | pH 4.7, 37°C, recombinant V6 mutant I84V | Human immunodeficiency virus 1 | |
0.000017 | - |
nelfinavir | pH 4.7, 37°C, recombinant V6 wild-type enzyme | Human immunodeficiency virus 1 | |
0.000023 | - |
nelfinavir | pH 4.7, 37°C, recombinant V6 mutant I54V | Human immunodeficiency virus 1 | |
0.000024 | - |
nelfinavir | pH 4.7, 37°C, recombinant V6 mutant I84V | Human immunodeficiency virus 1 | |
0.0000345 | - |
ritonavir | pH 4.7, 37°C, recombinant V6 mutant M46I/I84V | Human immunodeficiency virus 1 | |
0.000052 | - |
nelfinavir | pH 4.7, 37°C, recombinant V6 mutant M46I/I84V | Human immunodeficiency virus 1 | |
0.000062 | - |
indinavir | pH 4.7, 37°C, recombinant V6 mutant M46I/I84V | Human immunodeficiency virus 1 | |
0.0000624 | - |
ritonavir | pH 4.7, 37°C, recombinant V6 mutant M46I/I54V/I84V | Human immunodeficiency virus 1 | |
0.000069 | - |
indinavir | pH 4.7, 37°C, recombinant V6 wild-type enzyme | Human immunodeficiency virus 1 | |
0.0000932 | - |
ritonavir | pH 4.7, 37°C, recombinant V6 mutant I54V/I84V | Human immunodeficiency virus 1 | |
0.000116 | - |
nelfinavir | pH 4.7, 37°C, recombinant V6 mutant M46I | Human immunodeficiency virus 1 | |
0.000289 | - |
indinavir | pH 4.7, 37°C, recombinant V6 mutant I84V | Human immunodeficiency virus 1 | |
0.000398 | - |
indinavir | pH 4.7, 37°C, recombinant V6 mutant I54V | Human immunodeficiency virus 1 | |
0.000563 | - |
nelfinavir | pH 4.7, 37°C, recombinant V6 mutant M46I/I54V/I84V | Human immunodeficiency virus 1 | |
0.001259 | - |
nelfinavir | pH 4.7, 37°C, recombinant V6 mutant I54V/I84V | Human immunodeficiency virus 1 | |
0.001963 | - |
indinavir | pH 4.7, 37°C, recombinant V6 mutant M46I | Human immunodeficiency virus 1 | |
0.002914 | - |
indinavir | pH 4.7, 37°C, recombinant V6 mutant M46I/I54V/I84V | Human immunodeficiency virus 1 | |
0.004235 | - |
indinavir | pH 4.7, 37°C, recombinant V6 mutant I54V/I84V | Human immunodeficiency virus 1 |