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Literature summary for 3.4.21.97 extracted from

  • Batra, R.; Khayat, R.; Tong, L.
    Molecular mechanism for dimerization to regulate the catalytic activity of human cytomegalovirus protease (2001), Nat. Struct. Biol., 8, 810-817.
    View publication on PubMed
Search for more literature for 3.4.21.97

Cloned(Commentary)

Cloned (Comment) Organism
mutant enzymes expressed in Escherichia coli Human betaherpesvirus 5

Crystallization (Commentary)

Crystallization (Comment) Organism
sitting-drop vapour diffusion method, crystals of the S225Y mutant in complex with the peptidomimetic inhibitor BILC408 Human betaherpesvirus 5

Protein Variants

Protein Variants Comment Organism
A143Q all enzyme samples used in this experiments contain an additional mutation, A143Q. The mutation disables one of the internal cleavage sites but has little effects on the kinetic properties of the enzyme Human betaherpesvirus 5
A143Q/D217N the ratio of turnover number to Km-value is 11% of that for mutant enzyme A143Q Human betaherpesvirus 5
A143Q/D227N the ratio of turnover number to Km-value is 1273fold lower than that for mutant A143Q Human betaherpesvirus 5
A143Q/L229R mutant enzyme A143Q/L229R shows approximately a 20% decrease in helical content relative to the mutant enzyme A143Q, mutant enzyme A143Q/L229R has significant lower thermal stability than the mutant enzyme A143Q Human betaherpesvirus 5
A143Q/L229R mutation has little effect on the stability of the dimer, significant lower thermal stability than the mutant enzyme A143Q. The ratio of turnover number to Km-value is 1826fold lower than that for mutant enzyme A143Q Human betaherpesvirus 5
A143Q/R232H mutant enzyme A143Q/R232H essentially maintains wild-type catalytic activity, shows little change in the CD spectra compared to mutant A143Q enzyme, thermal stability is similar to that of mutant A143Q enzyme. The ratio of turnover number to Km-value is 5.1fold lower than that for mutant enzyme A143Q Human betaherpesvirus 5
A143Q/S225Y mutant enzyme with large conformational changes with 40% lower helical content relative to the mutant enzyme A143Q, mutant dimer has similar stability to the mutant A143Q dimer. Many of the conformational differences between the mutant A143Q and the mutant enzyme A143Q/S225Y are likely the direct result of the mutation itself. alphaF helix tilts away from the wild type dimer two-fold axis in the S225Y mutant, creating a space near the two-fold axis to accomodate the new Tyr side chain. The S225Y mutation is the likely trigger for the change in the monomer conformation and the dimer organization, significant lower thermal stability than the mutant enzyme A143Q. The ratio of turnover number to Km-value is 1273fold lower than that for mutant enzyme A143Q Human betaherpesvirus 5
A143Q/S225Y/L229R mutation has little effect on the stability of the dimer, mutant enzyme with large conformational changes with 40% lower helical content relative to the mutant enzyme A143Q, significant lower thermal stability than the mutant enzyme A143Q. The ratio of turnover number to Km-value is 1680fold lower than that for mutant enzyme A143Q Human betaherpesvirus 5

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0042
-
 4-[[4'-(dimethylamino)phenyl]azo]-benzoyl-Arg-Gly-Val-Val-Asn-Ala-Ser-Ser-Arg-Leu-Ala-5-[(2-'aminoethyl)-amino]naphthalenesulfonic acid pH 7.2, 30°C, mutant enzyme A143Q/R232H Human betaherpesvirus 5
0.0051
-
 4-[[4'-(dimethylamino)phenyl]azo]-benzoyl-Arg-Gly-Val-Val-Asn-Ala-Ser-Ser-Arg-Leu-Ala-5-[(2-'aminoethyl)-amino]naphthalenesulfonic acid pH 7.2, 30°C, mutant enzyme A143Q/D217N Human betaherpesvirus 5
0.0078
-
 4-[[4'-(dimethylamino)phenyl]azo]-benzoyl-Arg-Gly-Val-Val-Asn-Ala-Ser-Ser-Arg-Leu-Ala-5-[(2-'aminoethyl)-amino]naphthalenesulfonic acid pH 7.2, 30°C, mutant enzyme A143Q/D227N Human betaherpesvirus 5
0.0107
-
 4-[[4'-(dimethylamino)phenyl]azo]-benzoyl-Arg-Gly-Val-Val-Asn-Ala-Ser-Ser-Arg-Leu-Ala-5-[(2-'aminoethyl)-amino]naphthalenesulfonic acid pH 7.2, 30°C, mutant enzyme A143Q Human betaherpesvirus 5
0.012
-
 4-[[4'-(dimethylamino)phenyl]azo]-benzoyl-Arg-Gly-Val-Val-Asn-Ala-Ser-Ser-Arg-Leu-Ala-5-[(2-'aminoethyl)-amino]naphthalenesulfonic acid pH 7.2, 30°C, mutant enzyme A143Q/L229R Human betaherpesvirus 5
0.0136
-
 4-[[4'-(dimethylamino)phenyl]azo]-benzoyl-Arg-Gly-Val-Val-Asn-Ala-Ser-Ser-Arg-Leu-Ala-5-[(2-'aminoethyl)-amino]naphthalenesulfonic acid pH 7.2, 30°C, mutant enzyme A143Q/S225Y/L229R Human betaherpesvirus 5
0.0339
-
 4-[[4'-(dimethylamino)phenyl]azo]-benzoyl-Arg-Gly-Val-Val-Asn-Ala-Ser-Ser-Arg-Leu-Ala-5-[(2-'aminoethyl)-amino]naphthalenesulfonic acid pH 7.2, 30°C, mutant enzyme A143Q/S225Y Human betaherpesvirus 5

Organism

Organism UniProt Comment Textmining
Human betaherpesvirus 5
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-[[4'-(dimethylamino)phenyl]azo]-benzoyl-Arg-Gly-Val-Val-Asn-Ala-Ser-Ser-Arg-Leu-Ala-5-[(2'-aminoethyl)-amino]naphthalenesulfonic acid + H2O
-
 Human betaherpesvirus 5 4-[[4'-(dimethylamino)phenyl]azo]-benzoyl-Arg-Gly-Val-Val-Asn-Ala + Ser-Ser-Arg-Leu-Ala-5-[(2'-aminoethyl)-amino]naphthalenesulfonic acid
-
 ?

Subunits

Subunits Comment Organism
dimer dimerization is required for catalytic activity, appropriate dimer formation may be required to indirectly stabilize the protease oxyanion hole revealing a novel mechanism for dimerization to regulate enzyme activity Human betaherpesvirus 5

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
28
-
 10 min 50% loss of activity of mutant enzyme A143Q/S225Y Human betaherpesvirus 5
29
-
 10 min 50% loss of activity of mutant enzyme A143Q/L229R Human betaherpesvirus 5
30
-
 10 min 50% loss of activity of mutant enzyme A143Q/D227N and A143Q/S225Y/L229R Human betaherpesvirus 5
35
-
 10 min, 50% loss of activity of mutant enzymes A143Q and A143Q/R232H Human betaherpesvirus 5

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.000026
-
 4-[[4'-(dimethylamino)phenyl]azo]-benzoyl-Arg-Gly-Val-Val-Asn-Ala-Ser-Ser-Arg-Leu-Ala-5-[(2'-aminoethyl)-amino]naphthalenesulfonic acid pH 7.2, 30°C, mutant enzyme A143Q/D227N Human betaherpesvirus 5
0.000027
-
 4-[[4'-(dimethylamino)phenyl]azo]-benzoyl-Arg-Gly-Val-Val-Asn-Ala-Ser-Ser-Arg-Leu-Ala-5-[(2'-aminoethyl)-amino]naphthalenesulfonic acid pH 7.2, 30°C, mutant enzyme A143Q/L229R Human betaherpesvirus 5
0.000034
-
 4-[[4'-(dimethylamino)phenyl]azo]-benzoyl-Arg-Gly-Val-Val-Asn-Ala-Ser-Ser-Arg-Leu-Ala-5-[(2'-aminoethyl)-amino]naphthalenesulfonic acid pH 7.2, 30°C, mutant enzyme A143Q/S225Y/L229R Human betaherpesvirus 5
0.00092
-
 4-[[4'-(dimethylamino)phenyl]azo]-benzoyl-Arg-Gly-Val-Val-Asn-Ala-Ser-Ser-Arg-Leu-Ala-5-[(2'-aminoethyl)-amino]naphthalenesulfonic acid pH 7.2, 30°C, mutant enzyme A143Q/S225Y Human betaherpesvirus 5
0.0024
-
 4-[[4'-(dimethylamino)phenyl]azo]-benzoyl-Arg-Gly-Val-Val-Asn-Ala-Ser-Ser-Arg-Leu-Ala-5-[(2'-aminoethyl)-amino]naphthalenesulfonic acid pH 7.2, 30°C, mutant enzyme A143Q/D217N Human betaherpesvirus 5
0.00348
-
 4-[[4'-(dimethylamino)phenyl]azo]-benzoyl-Arg-Gly-Val-Val-Asn-Ala-Ser-Ser-Arg-Leu-Ala-5-[(2'-aminoethyl)-amino]naphthalenesulfonic acid pH 7.2, 30°C, mutant enzyme A143Q/R232H Human betaherpesvirus 5
0.046
-
 4-[[4'-(dimethylamino)phenyl]azo]-benzoyl-Arg-Gly-Val-Val-Asn-Ala-Ser-Ser-Arg-Leu-Ala-5-[(2'-aminoethyl)-amino]naphthalenesulfonic acid pH 7.2, 30°C, mutant enzyme A143Q Human betaherpesvirus 5