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Literature summary for 3.4.11.1 extracted from

  • Lee, Y.R.; Na, B.K.; Moon, E.K.; Song, S.M.; Joo, S.Y.; Kong, H.H.; Goo, Y.K.; Chung, D.I.; Hong, Y.
    Essential role for an M17 leucine aminopeptidase in encystation of Acanthamoeba castellanii (2015), PLoS ONE, 10, e0129884 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.11.1

Cloned(Commentary)

Cloned (Comment) Organism
gene LAP, sequence comparions and phylogenetic analysis, recombinant expression of His6-tagged TF-fused AcLAPr in Escherichia coli strain BL21, recombinant expression of EGFP-tagged enzyme in Acanthamoeba castellanii trophozoites Acanthamoeba castellanii

Protein Variants

Protein Variants Comment Organism
additional information AcLAP knockout using siRNA, AcLAP knockdown affects encystation of Acanthamoeba castellanii Acanthamoeba castellanii

Inhibitors

Inhibitors Comment Organism Structure
1,10-phenanthroline abolishes enzyme activity at 10 mM Acanthamoeba castellanii
bestatin inhibits enzyme activity by 77% at 0.1 mM Acanthamoeba castellanii
Co2+ activates at 0.1-1.0 mM, inhibitory at 1.0-10 mM Acanthamoeba castellanii
EDTA abolishes enzyme activity at 1 mM Acanthamoeba castellanii
Mn2+ activates at 0.1-1.0 mM, inhibitory at 1.0-10 mM Acanthamoeba castellanii
Ni2+ activates at 0.1-1.0 mM, inhibitory at 1.0-10 mM Acanthamoeba castellanii
Zn2+ activates at 0.1-1.0 mM, inhibitory at 1.0-10 mM Acanthamoeba castellanii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0038
-
 L-methionyl-7-amido-4-methylcoumarin pH 8.0, 37°C, recombinant enzyme Acanthamoeba castellanii
0.0135
-
 L-leucyl-7-amido-4-methylcoumarin pH 8.0, 37°C, recombinant enzyme Acanthamoeba castellanii
0.0414
-
 L-arginyl-7-amido-4-methylcoumarin pH 8.0, 37°C, recombinant enzyme Acanthamoeba castellanii

Localization

Localization Comment Organism GeneOntology No. Textmining
cytoplasm AcLAP is mainly localized in the cytoplasm of Acanthamoeba castellanii Acanthamoeba castellanii 5737
-

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ activates at 0.1-1.0 mM, inhibitory at 1.0-10 mM Acanthamoeba castellanii
Mg2+ activates at 0.1-10 mM Acanthamoeba castellanii
Mn2+ activates at 0.1-1.0 mM, inhibitory at 1.0-10 mM Acanthamoeba castellanii
additional information AcLAPr activity is progressively enhanced by Ni2+, Mn2+, Mg2+, Zn2+, and Co2+ but not by Ca2+. Among the divalent metal ions tested, Ni2+ (0.1-1.0 mM) increases AcLAPr activity the most by 5.5fold. Higher concentrations of all metal ions of 10 mM, except of Mg2+, inhibit AcLAPr activity Acanthamoeba castellanii
Ni2+ activates at 0.1-1.0 mM, inhibitory at 1.0-10 mM Acanthamoeba castellanii
Zn2+ activates at 0.1-1.0 mM, inhibitory at 1.0-10 mM Acanthamoeba castellanii

Organism

Organism UniProt Comment Textmining
Acanthamoeba castellanii A0A060A630
-
-

Source Tissue

Source Tissue Comment Organism Textmining
cyst AcLAP is highly expressed at a late stage of encystation, an approximate twofold increase is observed at 72 h post-encystation Acanthamoeba castellanii
-
trophozoite the expression of AcLAP in trophozoites shows no significant change of its level during 24 h and 48 h Acanthamoeba castellanii
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-arginyl-7-amido-4-methylcoumarin + H2O low activity Acanthamoeba castellanii L-arginine + 7-amino-4-methylcoumarin
-
 ?
L-leucyl-7-amido-4-methylcoumarin + H2O moderate activity Acanthamoeba castellanii L-leucine + 7-amino-4-methylcoumarin
-
 ?
L-methionyl-7-amido-4-methylcoumarin + H2O high activity Acanthamoeba castellanii L-methionine + 7-amino-4-methylcoumarin
-
 ?

Subunits

Subunits Comment Organism
? x * 61800, about, sequence calculation Acanthamoeba castellanii

Synonyms

Synonyms Comment Organism
AcLAP
-
 Acanthamoeba castellanii
M17 leucine aminopeptidase
-
 Acanthamoeba castellanii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Acanthamoeba castellanii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.13
-
 L-arginyl-7-amido-4-methylcoumarin pH 8.0, 37°C, recombinant enzyme Acanthamoeba castellanii
0.33
-
 L-methionyl-7-amido-4-methylcoumarin pH 8.0, 37°C, recombinant enzyme Acanthamoeba castellanii
0.83
-
 L-leucyl-7-amido-4-methylcoumarin pH 8.0, 37°C, recombinant enzyme Acanthamoeba castellanii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Acanthamoeba castellanii

General Information

General Information Comment Organism
evolution phylogenetic analysis reveals that AcLAP clusters into a clade closely related with LAPs from Amoebozoa, including Dictyostelium sp.. The enzyme contains the highly conserved M17 aminopeptidase family signature sequence (NTDAEGRL, residues 425-432) and highly conserved amino acid residues for metal binding (D350, D368, D427, E429) and catalytic site formation (K357, R431). The enzyme belongs to the M17 family of proteases Acanthamoeba castellanii
malfunction lack or downregulation of AcLAP activity causes ultrastructural changes of the cell wall in Acanthamoeba castellanii that are closely associated with inhibition of cyst formation Acanthamoeba castellanii
physiological function AcLAP is a typical M17 family enzyme that plays an essential role during encystation of Acanthamoeba castellanii Acanthamoeba castellanii

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.14
-
 L-arginyl-7-amido-4-methylcoumarin pH 8.0, 37°C, recombinant enzyme Acanthamoeba castellanii
61.5
-
 L-leucyl-7-amido-4-methylcoumarin pH 8.0, 37°C, recombinant enzyme Acanthamoeba castellanii
86.8
-
 L-methionyl-7-amido-4-methylcoumarin pH 8.0, 37°C, recombinant enzyme Acanthamoeba castellanii