3.4.23.47: HIV-2 retropepsin

inactive mutant forms a dimer

DELTA96-99

dimer interface mutations such as a deletion of the C-terminal residues 96-99 (PR21-95), drastically increase the Kd. Deletion mutant consists predominantly of folded monomers. Addition of 2fold excess active-site inhibitor does not promote dimerization

E37K

mutation significantly retards autoproteolytic cleavage during expression. Mutant shows a higher dimer dissociation constant, Kd, compared to wild-type. Km and kcat values for substrate Lys-Ala-Arg-Val-Nle-(4-nitrophenylalanine)-Glu-Ala-Nle comparable to wild-type

I32V

site-directed mutagenesis, mutation analogous to wild-type HIV-1 protease sequence (EC 3.4.23.16). The mutation increases the susceptibility of HIV-2 to multiple protease inhibitors

I50V

IC50 (mM): value above 10 (amprenavir), 0.000072 (idinavir), 0.000132 (nelfinavir), 0.000316 (tipranavir), 0.000203 (lopinavir)

I54M

2 entries

I54M/I84V

IC50 (mM): value above 10 (amprenavir), 0.000388 (idinavir), 0.001747 (nelfinavir), 0.002028 (tipranavir), 0.000375 (lopinavir)

I54M/L90M

IC50 (mM): value above 10 (amprenavir), 0.001112 (idinavir), 0.002564 (nelfinavir), 0.000145 (tipranavir), 0.000524 (lopinavir)

I54M/L99F

IC50 (mM): value above 10 (amprenavir), 0.000825 (idinavir), 0.002241 (nelfinavir), 0.002908 (tipranavir), 0.000414 (lopinavir)

I82F

IC50 (mM): 0.000965 (amprenavir), 0.000544 (idinavir), 0.000244 (nelfinavir), 0.000306 (tipranavir), 0.000145 (lopinavir)

I82L

IC50 (mM): 0.000528 (amprenavir), 0.000064 (idinavir), 0.000106 (nelfinavir), 0.00016 (tipranavir), 0.000075 (lopinavir)

I82L/L99F

IC50 (mM): 0.000098 (amprenavir), 0.000143 (idinavir), 0.000961 (nelfinavir), 0.000524 (tipranavir), 0.000496 (lopinavir)

I82V

site-directed mutagenesis, mutation analogous to wild-type HIV-1 protease sequence (EC 3.4.23.16)

K45R

mutant is resistant toward HIV-2 protease inhibitor lopinavir and saquinavir in yeast

K57L

site-directed mutagenesis, the mutation is experimentally introduced to help the crystallographic process

754210

L90M

IC50: 0.0125 (lopinavir), 0.554 (saquinavir), L90M mutation present in primary HIV-2 isolate, modifies the HIV-2 protease susceptibility to saquinavir but not lopinavir

L99F

IC50: 0.0111 (lopinavir), 0.1472 (saquinavir)

M76L

site-directed mutagenesis, mutation analogous to wild-type HIV-1 protease sequence (EC 3.4.23.16). The mutation increases the susceptibility of HIV-2 to multiple protease inhibitors

M76V

mutant is resistant toward HIV-2 protease inhibitor lopinavir and saquinavir in yeast

T26A

dimer interface mutations in the active site of PR2 drastically increase the Kd. T26A consists predominantly of folded monomers. Addition of 2fold excess active-site inhibitor promotes dimerization of mutant PR2T26A

V47A/D17N

passage of HIV-2MS with increasing concentrations of lopinavir selects mutations V47A and D17N in the HIV-2 protease gene. Introduction of both 17N and 47A either individually or together into HIV-2ROD molecular infectious clones shows that the single V47A substitution in HIV-2 results in a substantial reduction in susceptibility to lopinavir. This mutant retains wild-type susceptibility to other proteinase inhibitors and is hypersusceptible to atazanavir and saquinavir

677593

V47I

site-directed mutagenesis, mutation analogous to wild-type HIV-1 protease sequence (EC 3.4.23.16). The mutation increases the susceptibility of HIV-2 to multiple protease inhibitors

V47I/M76L/I32V/I82V

site-directed mutagenesis, mutation analogous to wild-type HIV-1 protease sequence (EC 3.4.23.16). The combination of the four amino acid changes in HIV-2 protease confers a pattern of protease inhibitor susceptibility comparable to that of HIV-1

V71I, L90M, I89V