Literature summary for 3.4.23.16 extracted from

Clemente, J.C.; Moose, R.E.; Hemrajani, R.; Whitford, L.R.; Govindasamy, L.; Reutzel, R.; McKenna, R.; Agbandje-McKenna, M.; Goodenow, M.M.; Dunn, B.M.
Comparing the accumulation of active- and nonactive-site mutations in the HIV-1 protease (2004), Biochemistry, 43, 12141-12151.

Search for more literature for 3.4.23.16

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 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

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

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

Inhibitors	Comment	Organism
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 [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	kinetics, LAI wild-type enzyme	Human immunodeficiency virus 1

Organism

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

Purification (Commentary)

Purification (Comment)	Organism
refolded recombinant wild-type and mutant enzymes by ammonium sulfate precipitation and gel filtration	Human immunodeficiency virus 1

Renatured (Commentary)

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 and Products (Substrate)

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

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

Temperature Optimum [°C]

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

pH Optimum

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

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism
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