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 refolded recombinant wild-type and mutant enzymes in complex with inhibitors, hanging drop vapour diffusion method, room temperature, preincubation of 1.8-3.5 mg/ml protein with inhibitor in a ratio of 1:5 - 1:20, for mutant L24I: the reservoir solution contains 0.2 M sodium phosphate, pH 5.0-6.0, 10% v/v DMSO, and 20-40% saturated ammonium sulfate, for mutants I50V and G73S: the reservoir solution contains 0.1 M sodium citrate/0.2 M sodium phosphate, pH 5.4-5.8, 8-13% DMSO and 6-20% saturated ammonium sulfate, protein and reservoir solutions are mixed in ratio of 1:1, X-ray diffraction structure determination snd analysis at 1.1-1.5 A resolution | Human immunodeficiency virus 1 |
Protein Variants | Comment | Organism |
---|---|---|
G73S | site-directed mutagenesis, drug-resistant mutant | Human immunodeficiency virus 1 |
I50V | site-directed mutagenesis, drug-resistant mutant | Human immunodeficiency virus 1 |
L42I | site-directed mutagenesis, drug-resistant mutant | Human immunodeficiency virus 1 |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ace-T-I-Nle-r-Nle-Q-R | substrate analogue peptide inhibitor | Human immunodeficiency virus 1 | |
indinavir | - |
Human immunodeficiency virus 1 | |
R-V-L-r-F-E-A-Nle | substrate analogue peptide inhibitor | Human immunodeficiency virus 1 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | substrate specificity and kinetics of wild-type and mutant enzymes, overview | Human immunodeficiency virus 1 | |
0.046 | - |
Lys-Ala-Arg-Val-Nle-4-nitro-Phe-Glu-Ala-Nle-amide | pH 5.0, 25°C, mutant G73S | Human immunodeficiency virus 1 | |
0.31 | - |
Lys-Ala-Arg-Val-Nle-4-nitro-Phe-Glu-Ala-Nle-amide | pH 5.0, 25°C, mutant L24I | Human immunodeficiency virus 1 | |
0.5 | - |
Lys-Ala-Arg-Val-Nle-4-nitro-Phe-Glu-Ala-Nle-amide | pH 5.0, 25°C, mutant I50V | Human immunodeficiency virus 1 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
Gag polyprotein + H2O | Human immunodeficiency virus 1 | - |
? | - |
? | |
Gag-Pol polyprotein + H2O | Human immunodeficiency virus 1 | - |
? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Human immunodeficiency virus 1 | - |
i.e. HIV-1 | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from inclusion bodies after expression in Escherichia coli strain BL21(DE3) | Human immunodeficiency virus 1 |
Renatured (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from inclusion bodies after expression in Escherichia coli strain BL21(DE3), solubilization by urea and refolding | Human immunodeficiency virus 1 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
Gag polyprotein + H2O | - |
Human immunodeficiency virus 1 | ? | - |
? | |
Gag polyprotein + H2O | CA/p2 cleavage site is KARVL*AEAMS | Human immunodeficiency virus 1 | ? | - |
? | |
Gag-Pol polyprotein + H2O | - |
Human immunodeficiency virus 1 | ? | - |
? | |
Gag-Pol polyprotein + H2O | NC/p1 cleavage site is ERQAN*FLGKI | Human immunodeficiency virus 1 | ? | - |
? | |
Lys-Ala-Arg-Val-Nle-4-nitro-Phe-Glu-Ala-Nle-amide + H2O | the peptide substrate mimicks the HIV-1 CA/p2 cleavage site | Human immunodeficiency virus 1 | Lys-Ala-Arg-Val-Nle + 4-nitro-Phe-Glu-Ala-Nle-amide | - |
? | |
additional information | substrate specificity and kinetics of wild-type and mutant enzymes, overview | Human immunodeficiency virus 1 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | - |
Human immunodeficiency virus 1 |
Synonyms | Comment | Organism |
---|---|---|
HIV-1 protease | - |
Human immunodeficiency virus 1 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Human immunodeficiency virus 1 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
61 | - |
Lys-Ala-Arg-Val-Nle-4-nitro-Phe-Glu-Ala-Nle-amide | pH 5.0, 25°C, mutant L24I | Human immunodeficiency virus 1 | |
280 | - |
Lys-Ala-Arg-Val-Nle-4-nitro-Phe-Glu-Ala-Nle-amide | pH 5.0, 25°C, mutant G73S | Human immunodeficiency virus 1 | |
480 | - |
Lys-Ala-Arg-Val-Nle-4-nitro-Phe-Glu-Ala-Nle-amide | pH 5.0, 25°C, mutant I50V | Human immunodeficiency virus 1 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | - |
assay at | Human immunodeficiency virus 1 |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00000054 | - |
indinavir | pH 5.0, 25°C, wild-type enzyme | Human immunodeficiency virus 1 | |
0.00000055 | - |
indinavir | pH 5.0, 25°C, mutant G73S | Human immunodeficiency virus 1 | |
0.0000014 | - |
indinavir | pH 5.0, 25°C, mutant L24I | Human immunodeficiency virus 1 | |
0.000002 | - |
Ace-T-I-Nle-r-Nle-Q-R | pH 5.0, 25°C, mutant L24I | Human immunodeficiency virus 1 | |
0.0000022 | - |
Ace-T-I-Nle-r-Nle-Q-R | pH 5.0, 25°C, wild-type enzyme | Human immunodeficiency virus 1 | |
0.0000033 | - |
Ace-T-I-Nle-r-Nle-Q-R | pH 5.0, 25°C, mutant G73S | Human immunodeficiency virus 1 | |
0.0000035 | - |
R-V-L-r-F-E-A-Nle | pH 5.0, 25°C, mutant L24I | Human immunodeficiency virus 1 | |
0.000027 | - |
indinavir | pH 5.0, 25°C, mutant I50V | Human immunodeficiency virus 1 | |
0.000041 | - |
Ace-T-I-Nle-r-Nle-Q-R | pH 5.0, 25°C, mutant I50V | Human immunodeficiency virus 1 | |
0.000075 | - |
R-V-L-r-F-E-A-Nle | pH 5.0, 25°C, wild-type enzyme | Human immunodeficiency virus 1 | |
0.00023 | - |
R-V-L-r-F-E-A-Nle | pH 5.0, 25°C, mutant I50V | Human immunodeficiency virus 1 | |
0.00033 | - |
R-V-L-r-F-E-A-Nle | pH 5.0, 25°C, mutant G73S | Human immunodeficiency virus 1 |