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Literature summary for 3.4.23.16 extracted from

  • Liu, F.; Boross, P.I.; Wang, Y.F.; Tozser, J.; Louis, J.M.; Harrison, R.W.; Weber, I.T.
    Kinetic, stability, and structural changes in high-resolution crystal structures of HIV-1 protease with drug-resistant mutations L24I, I50V, and G73S (2005), J. Mol. Biol., 354, 789-800.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) in inclusion bodies Human immunodeficiency virus 1

Crystallization (Commentary)

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

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

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 [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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Human immunodeficiency virus 1
-
i.e. HIV-1
-

Purification (Commentary)

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 (Commentary)

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 and Products (Substrate)

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

Subunits Comment Organism
dimer
-
Human immunodeficiency virus 1

Synonyms

Synonyms Comment Organism
HIV-1 protease
-
Human immunodeficiency virus 1

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Human immunodeficiency virus 1

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
5
-
assay at Human immunodeficiency virus 1

Ki Value [mM]

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