Cloned (Comment) | Organism |
---|---|
HIV-1 protease clone for expression in Escherichia coli is constructed with the substitutions Q7K/L33I/L63I/C67A/C95A. The expressed enzyme is purified from inclusion bodies | Human immunodeficiency virus 1 |
Crystallization (Comment) | Organism |
---|---|
mutant enzymes R8Q, K45I and L90M are crystallized with the inhibitor RVL-(reduced peptide bond)-FEA-Nle-NH2 | Human immunodeficiency virus 1 |
Protein Variants | Comment | Organism |
---|---|---|
D30N | similar to wild-type enzyme in stability towards urea denaturation. The kcat/Km ratio for the substrate KARVLAEAMS is 113% of that for the autoproteolysis resistant variant Q7K/L33I/L63I/C67A/C95A | Human immunodeficiency virus 1 |
G48V | the kcat/Km ratio for the substrate KARVLAEAMS is 55% of that for the autoproteolysis resistant variant Q7K/L33I/L63I/C67A/C95A | Human immunodeficiency virus 1 |
G48V | half maximal activity at 0.7 M urea compared to 1.8 M for the wild-type enzyme | Human immunodeficiency virus 1 |
K45I | mutant with significantly increased stability with half-maximal activity at 3.3 mM urea compared to 1.8 M urea for the wild-type enzyme. The kcat/Km ratio for the substrate KARVLAEAMS is 106% of that for the autoproteolysis resistant variant Q7K/L33I/L63I/C67A/C95A | Human immunodeficiency virus 1 |
L90M | the kcat/Km ratio for the substrate KARVLAEAMS is 44% of that for the autoproteolysis resistant variant Q7K/L33I/L63I/C67A/C95A. Half maximal activity at 1.0 M urea compared to 1.8 M for the wild-type enzyme | Human immunodeficiency virus 1 |
M46L | the kcat/Km ratio for the substrate KARVLAEAMS is 63% of that for the autoproteolysis resistant variant Q7K/L33I/L63I/C67A/C95A | Human immunodeficiency virus 1 |
N88D | mutant with significantly increased stability with half-maximal activity at 3.1 mM urea compared to 1.8 M urea for the wild-type enzyme. The kcat/Km ratio for the substrate KARVLAEAMS is 39% of that for the autoproteolysis resistant variant Q7K/L33I/L63I/C67A/C95A | Human immunodeficiency virus 1 |
Q7K/L33I/L63I/C67A/C95A | the kinetic parameters are nearly identical to those of the native enzyme. Half maximal activity at 1.9 M urea compared to 1.8 M for the wild-type enzyme | Human immunodeficiency virus 1 |
Q7K/L33I/L63I/C67A/C95A | mutant enzyme with restricted autoproteolysis, self-degradation | Human immunodeficiency virus 1 |
R8Q | half maximal activity at 1.3 M urea compared to 1.8 M for the wild-type enzyme. The kcat/Km ratio for the substrate KARVLAEAMS is 15% of that for the autoproteolysis resistant variant Q7K/L33I/L63I/C67A/C95A | Human immunodeficiency virus 1 |
V82S | similar to wild-type enzyme in stability towards urea denaturation. The kcat/Km ratio for the substrate KARVLAEAMS is 24% of that for the autoproteolysis resistant variant Q7K/L33I/L63I/C67A/C95A | Human immunodeficiency virus 1 |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | RVL-(reduced peptide bond)-FEA-Nle-NH2 and Ac-TI-Nle-(reduced peptide bond)-Nle-QR-NH2 | Human immunodeficiency virus 1 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.06 | - |
KARV-Nle-Phe(NO2)-EA-Nle | pH 5.0, wild-type enzyme | Human immunodeficiency virus 1 | |
0.075 | - |
KARV-Nle-Phe(NO2)-EA-Nle | pH 5.0, mutant enzyme Q7K/L33I/L63I/C67A/C95A | Human immunodeficiency virus 1 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Human immunodeficiency virus 1 | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
KARV-Nle-Phe(NO2)-EA-Nle + H2O | - |
Human immunodeficiency virus 1 | KARV-Nle + Phe(NO2)-EA-Nle | - |
? | |
KARV-Nle-Phe(NO2)-EA-Nle-NH2 + H2O | - |
Human immunodeficiency virus 1 | KARV-Nle + Phe(NO2)-EA-Nle-NH2 | - |
? | |
KARVLAEAMS + H2O | - |
Human immunodeficiency virus 1 | KARVL + AEAMS | - |
? | |
TLNFPISPKK + H2O | - |
Human immunodeficiency virus 1 | TLNF + PISPKK | - |
? | |
VSFNFPQITKK + H2O | - |
Human immunodeficiency virus 1 | VSFNF + PQITKK | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.767 | - |
KARV-Nle-Phe(NO2)-EA-Nle | pH 5.0, mutant enzyme Q7K/L33I/L63I/C67A/C95A | Human immunodeficiency virus 1 | |
1.23 | - |
KARV-Nle-Phe(NO2)-EA-Nle | pH 5.0, wild-type enzyme | Human immunodeficiency virus 1 |