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Literature summary for 3.4.14.12 extracted from

  • Ito, K.; Nakajima, Y.; Xu, Y.; Yamada, N.; Onohara, Y.; Ito, T.; Matsubara, F.; Kabashima, T.; Nakayama, K.; Yoshimoto, T.
    Crystal structure and mechanism of tripeptidyl activity of prolyl tripeptidyl aminopeptidase from Porphyromonas gingivalis (2006), J. Mol. Biol., 362, 228-240.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli strain JM83 Porphyromonas gingivalis
into the pQE30 vector, lacking the region corresponding to residues 1-38, for transformation of Escherichia coli M15 cells Porphyromonas gingivalis

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging drop vapour diffusion method, with 1.1 M potassium/sodium tartrate, 100 mM Ches buffer (pH 9.0), and 200 mM lithium sulfate Porphyromonas gingivalis
the structure of the ligand-free enzyme is solved at 2.1 A resolution, the structure of the S603A mutant-substrate complex is solved at 2.9 A resolution Porphyromonas gingivalis

Protein Variants

Protein Variants Comment Organism
S603A inactive Porphyromonas gingivalis
S603A inactive mutant Porphyromonas gingivalis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.42
-
Gly-Ala-Pro-beta-naphthylamide
-
Porphyromonas gingivalis
0.42
-
Gly-Ala-Pro-2-naphthylamide in 20 mM Tris-HCl buffer (pH 7.0), at 37°C Porphyromonas gingivalis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
72500
-
2 * 72500, gel filtration Porphyromonas gingivalis
77000
-
determined by SDS-PAGE Porphyromonas gingivalis
145000
-
gel filtration Porphyromonas gingivalis
145000
-
determined by gel filtration Porphyromonas gingivalis
154000
-
SDS-PAGE Porphyromonas gingivalis

Organism

Organism UniProt Comment Textmining
Porphyromonas gingivalis Q7MUW6
-
-

Purification (Commentary)

Purification (Comment) Organism
Toyopearl HW65C column chromatography and DEAE-Toyopearl column chromatography Porphyromonas gingivalis
using a Toyopearl HW65C and a DEAE-Toyopearl column Porphyromonas gingivalis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.12
-
cell free extract Porphyromonas gingivalis
32.1
-
purification step, after protamine sulfate treatment Porphyromonas gingivalis
74.7
-
purification step, ammonium sulfate precipitation, 35-70% pellet Porphyromonas gingivalis
118
-
purification step, chromatography on a Toyopearl HW65C column Porphyromonas gingivalis
160
-
purification step, chromatography on a DEAE-Toyopearl column Porphyromonas gingivalis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Ala-Phe-Pro-2-naphthylamide + H2O
-
Porphyromonas gingivalis Ala-Phe-Pro + 2-naphthylamine
-
?
Ala-Phe-Pro-beta-naphthylamide + H2O
-
Porphyromonas gingivalis Ala-Phe-Pro + beta-naphthylamine
-
?
Gly-Ala-Pro-2-naphthylamide + H2O
-
Porphyromonas gingivalis Gly-Ala-Pro + 2-naphthylamine
-
?
Gly-Ala-Pro-beta-naphthylamide + H2O
-
Porphyromonas gingivalis Gly-Ala-Pro + beta-naphthylamine
-
?
additional information no activity with Pro-2-naphthylamide, Gly-Pro-2-naphthylamide, Z-Gly-Pro-2-naphthylamide, Z-Gly-Ala-Pro-2-naphthylamide, Z-Ala-Gly-Pro-2-naphthylamide, Gly-Ala-Gly-Pro-2-naphthylamide, Z-Gly-Ala-Gly-Pro-2-naphthylamide, and Ala-Gly-Ala-Gly-Pro-2-naphthylamide Porphyromonas gingivalis ?
-
?

Subunits

Subunits Comment Organism
homodimer
-
Porphyromonas gingivalis
homodimer 2 * 77000, SDS-PAGE Porphyromonas gingivalis
homodimer 2 * 72500, gel filtration Porphyromonas gingivalis

Synonyms

Synonyms Comment Organism
prolyl tripeptidyl aminopeptidase
-
Porphyromonas gingivalis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
42
-
-
Porphyromonas gingivalis
42
-
activity assay, Ala-Phe-Pro-beta-naphthylamide is used as a substrate Porphyromonas gingivalis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
354
-
Gly-Ala-Pro-beta-naphthylamide
-
Porphyromonas gingivalis
354
-
Gly-Ala-Pro-2-naphthylamide in 20 mM Tris-HCl buffer (pH 7.0), at 37°C Porphyromonas gingivalis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.2
-
-
Porphyromonas gingivalis
7.2
-
activity assay, Ala-Phe-Pro-beta-naphthylamide is used as a substrate Porphyromonas gingivalis