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Literature summary extracted from

  • Sarnovsky, R.; Rea, J.; Makowski, M.; Hertle, R.; Kelly, C.; Antignani, A.; Pastrana, D.V.; Fitzgerald, D.J.
    Proteolytic cleavage of a C-terminal prosequence, leading to autoprocessing at the N terminus, activates leucine aminopeptidase from Pseudomonas aeruginosa (2009), J. Biol. Chem., 284, 10243-10253.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.4.11.10 additional information full-length LAP is activated in a two-step process: proteolytic cleavage at the C terminus is followed by an intramolecular autocatalytic removal of a 12-amino acid propeptide at the N terminus Pseudomonas aeruginosa

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.11.10 expressed in Escherichia coli BL21-Star(DE3) cells Pseudomonas aeruginosa

Protein Variants

EC Number Protein Variants Comment Organism
3.4.11.10 D308A inactive Pseudomonas aeruginosa
3.4.11.10 D369A inactive Pseudomonas aeruginosa

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.4.11.10 Zn2+ 0.05 mM, required for activity Pseudomonas aeruginosa

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.4.11.10 50000
-
 active truncated recombinant enzyme, gel filtration Pseudomonas aeruginosa
3.4.11.10 54930
-
 recombinant enzyme, MALDI-TOF mass spectrometry Pseudomonas aeruginosa
3.4.11.10 66000
-
 inactive full-length recombinant enzyme, gel filtration Pseudomonas aeruginosa

Organism

EC Number Organism UniProt Comment Textmining
3.4.11.10 Pseudomonas aeruginosa
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.11.10 Q-Sepharose column chromatography and Superdex 200 gel filtration Pseudomonas aeruginosa

Renatured (Commentary)

EC Number Renatured (Comment) Organism
3.4.11.10 recombinant LAP is refolded by adding guanidine-solubilized protein to the following solution: 0.5 M L-Arg, 50 mM Tris, pH 8.0, 1 mM CaCl2, and 0.05 mM ZnCl2 Pseudomonas aeruginosa

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.11.10 Leu-4-nitroanilide + H2O
-
 Pseudomonas aeruginosa Leu + 4-nitroaniline
-
 ?
3.4.11.10 additional information LAP shows preference for leucine and methionine over other substrates Pseudomonas aeruginosa ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.4.11.10 LAP
-
 Pseudomonas aeruginosa
3.4.11.10 leucine aminopeptidase
-
 Pseudomonas aeruginosa
3.4.11.10 rLAP55 recombinant enzyme, full-length rLAP55 is enzymatically inactive Pseudomonas aeruginosa