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Literature summary for 3.4.11.21 extracted from

  • Zheng, J.; Cheng, Z.; Jia, H.; Zheng, Y.
    Characterization of aspartyl aminopeptidase from Toxoplasma gondii (2016), Sci. Rep., 6, 34448 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.11.21

Application

Application Comment Organism
drug development enzyme TgAAP deletion results in inhibition, but not abolition, of invasion and growth of Toxoplasma gondii in cell culture, thus indicating that the enzyme may be a promising adjunct target for anti-toxoplasma drug development Toxoplasma gondii

Cloned(Commentary)

Cloned (Comment) Organism
gene TGGT1_297970, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of GST-tagged enzyme in Escherichia coli Toxoplasma gondii

Protein Variants

Protein Variants Comment Organism
additional information knockout of enzyme TgAAP using the CRISPR/Cas9 knockout system, knockout plasmid pCD-TgAAP is constructed by inserting DHFR coding sequence and TgAAP-specific single guide RNA (gRNA) into plasmid pSAG1::CAS9-U6::sgUPRT, which contains Cas9, nuclear localization signal (NLS), and expresses GFP fusion element. The substrate activity is almost completely lost in the DELTATgAAP strain, thus, no other enzyme complements the TgAAP activity Toxoplasma gondii

Inhibitors

Inhibitors Comment Organism Structure
Ca2+ slight inhibition Toxoplasma gondii
CID 23724194 a PfM18AAP inhibitor, leads to over 50% inhibition at 1 mM Toxoplasma gondii
Cu2+ strong inhibition Toxoplasma gondii
Zn2+ strong inhibition Toxoplasma gondii

Localization

Localization Comment Organism GeneOntology No. Textmining
cytoplasm
-
 Toxoplasma gondii 5737
-

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ activates about 6fold at 1 mM Toxoplasma gondii
Fe2+ activates over 2fold at 1 mM Toxoplasma gondii
Mg2+ activates slightly at 1 mM Toxoplasma gondii
Mn2+ activates about 2.5fold at 1 mM Toxoplasma gondii
additional information AAP is a metalloaminopeptidase, M18 metalloaminopeptidases use metal ions to increase their substrate specificity for N-terminal acidic amino acids, beyond aspartic and glutamic acids Toxoplasma gondii
Ni2+ activates about 2fold at 1 mM Toxoplasma gondii

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
240000
-
 recombinant enzyme trimer, native PAGE Toxoplasma gondii
960000
-
 recombinant enzyme dodecamer, native PAGE Toxoplasma gondii

Organism

Organism UniProt Comment Textmining
Toxoplasma gondii S7UJV5
-
-
Toxoplasma gondii ATCC 50853 S7UJV5
-
-
Toxoplasma gondii GT1 S7UJV5
-
-
Toxoplasma gondii RH S7UJV5
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant GST-tagged enzyme from Escherichia coli by glutathione affinity chromatography and dialysis Toxoplasma gondii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-Asp-7-amido-4-methylcoumarin + H2O
-
 Toxoplasma gondii L-Asp + 7-amino-4-methylcoumarin
-
 ?
L-Asp-7-amido-4-methylcoumarin + H2O
-
 Toxoplasma gondii GT1 L-Asp + 7-amino-4-methylcoumarin
-
 ?
L-Asp-7-amido-4-methylcoumarin + H2O
-
 Toxoplasma gondii RH L-Asp + 7-amino-4-methylcoumarin
-
 ?
L-Asp-7-amido-4-methylcoumarin + H2O
-
 Toxoplasma gondii ATCC 50853 L-Asp + 7-amino-4-methylcoumarin
-
 ?
L-Glu-7-amido-4-methylcoumarin + H2O
-
 Toxoplasma gondii L-Glu + 7-amino-4-methylcoumarin
-
 ?
L-Glu-7-amido-4-methylcoumarin + H2O
-
 Toxoplasma gondii GT1 L-Glu + 7-amino-4-methylcoumarin
-
 ?
L-Glu-7-amido-4-methylcoumarin + H2O
-
 Toxoplasma gondii RH L-Glu + 7-amino-4-methylcoumarin
-
 ?
L-Glu-7-amido-4-methylcoumarin + H2O
-
 Toxoplasma gondii ATCC 50853 L-Glu + 7-amino-4-methylcoumarin
-
 ?
additional information rTgAAP has a strict preference for Asp-7-amido-4-methylcoumarin and Glu-7-amido-4-methylcoumarin, the catalytic efficiency of the enzyme is higher with L-Glu-7-amido-4-methylcoumarin than with L-Asp-7-amido-4-methylcoumarin. No activity with Ala-, Arg-, Cys-, Leu-, Lys, Met-, Phe, Trp-, and Tyr-7-amido-4-methylcoumarin, and poor activity with His-, Ser-, and Pro-7-amido-4-methylcoumarin Toxoplasma gondii ?
-
 ?
additional information rTgAAP has a strict preference for Asp-7-amido-4-methylcoumarin and Glu-7-amido-4-methylcoumarin, the catalytic efficiency of the enzyme is higher with L-Glu-7-amido-4-methylcoumarin than with L-Asp-7-amido-4-methylcoumarin. No activity with Ala-, Arg-, Cys-, Leu-, Lys, Met-, Phe, Trp-, and Tyr-7-amido-4-methylcoumarin, and poor activity with His-, Ser-, and Pro-7-amido-4-methylcoumarin Toxoplasma gondii GT1 ?
-
 ?
additional information rTgAAP has a strict preference for Asp-7-amido-4-methylcoumarin and Glu-7-amido-4-methylcoumarin, the catalytic efficiency of the enzyme is higher with L-Glu-7-amido-4-methylcoumarin than with L-Asp-7-amido-4-methylcoumarin. No activity with Ala-, Arg-, Cys-, Leu-, Lys, Met-, Phe, Trp-, and Tyr-7-amido-4-methylcoumarin, and poor activity with His-, Ser-, and Pro-7-amido-4-methylcoumarin Toxoplasma gondii RH ?
-
 ?
additional information rTgAAP has a strict preference for Asp-7-amido-4-methylcoumarin and Glu-7-amido-4-methylcoumarin, the catalytic efficiency of the enzyme is higher with L-Glu-7-amido-4-methylcoumarin than with L-Asp-7-amido-4-methylcoumarin. No activity with Ala-, Arg-, Cys-, Leu-, Lys, Met-, Phe, Trp-, and Tyr-7-amido-4-methylcoumarin, and poor activity with His-, Ser-, and Pro-7-amido-4-methylcoumarin Toxoplasma gondii ATCC 50853 ?
-
 ?

Subunits

Subunits Comment Organism
oligomer x * 80000, recombinant GST-tagged enzyme, SDS-PAGE, x * 56000, about, untagged enzyme, sequence calculation Toxoplasma gondii

Synonyms

Synonyms Comment Organism
AAP
-
 Toxoplasma gondii
aspartyl aminopeptidase
-
 Toxoplasma gondii
TgAAP
-
 Toxoplasma gondii
TGGT1_297970
-
 Toxoplasma gondii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Toxoplasma gondii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at, recombinant enzyme Toxoplasma gondii
8
-
 assay at, native enzyme Toxoplasma gondii

General Information

General Information Comment Organism
evolution enzyme AAP belongs to the M18 family of peptidases Toxoplasma gondii
malfunction TgAAP knockout inhibits the attachment/invasion, replication, and substrate-specific activity in Toxoplasma gondii. TgAAP knockout affects the growth of Toxoplasma gondii but does not completely abolish parasite replication and growth Toxoplasma gondii

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
85.66
-
 L-Asp-7-amido-4-methylcoumarin pH 7.5, 37°C, recombinant enzyme, with Co2+ Toxoplasma gondii
190.16
-
 L-Glu-7-amido-4-methylcoumarin pH 7.5, 37°C, recombinant enzyme, with Co2+ Toxoplasma gondii