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Literature summary extracted from

  • Gu, Y.Q.; Walling, L.L.
    Identification of residues critical for activity of the wound-induced leucine aminopeptidase (LAP-A) of tomato (2002), Eur. J. Biochem., 269, 1630-1640.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.11.1 overexpression of His-tagged wild-type and mutants in Escherichia coli Solanum lycopersicum

Protein Variants

EC Number Protein Variants Comment Organism
3.4.11.1 D347E oligonucleotide site-directed mutagenesis, formation of a more complex structure compared to the wild-type enzyme, nearly catalytically inactive with dipeptide substrates Solanum lycopersicum
3.4.11.1 D347G oligonucleotide site-directed mutagenesis, formation of a stable hexamer corresponding to the wild-type enzyme, nearly catalytically inactive with dipeptide substrates Solanum lycopersicum
3.4.11.1 D347I oligonucleotide site-directed mutagenesis, no formation of stable hexamer, inactive mutant Solanum lycopersicum
3.4.11.1 D347N oligonucleotide site-directed mutagenesis, no formation of stable hexamer, inactive mutant Solanum lycopersicum
3.4.11.1 D347R oligonucleotide site-directed mutagenesis, formation of a stable hexamer corresponding to the wild-type enzyme, reduced catalytic activity with dipeptide substrates compared to the wild-type Solanum lycopersicum
3.4.11.1 D347S oligonucleotide site-directed mutagenesis, no formation of stable hexamer, inactive mutant Solanum lycopersicum
3.4.11.1 D347V oligonucleotide site-directed mutagenesis, no formation of stable hexamer, inactive mutant Solanum lycopersicum
3.4.11.1 D347Y oligonucleotide site-directed mutagenesis, formation of a more complex structure compared to the wild-type enzyme Solanum lycopersicum
3.4.11.1 D354E oligonucleotide site-directed mutagenesis, no formation of stable hexamer, inactive mutant Solanum lycopersicum
3.4.11.1 D354M oligonucleotide site-directed mutagenesis, reduced catalytic activity with dipeptide substrates compared to the wild-type Solanum lycopersicum
3.4.11.1 D354R oligonucleotide site-directed mutagenesis, no formation of stable hexamer, reduced catalytic activity with dipeptide substrates compared to the wild-type Solanum lycopersicum
3.4.11.1 E429A oligonucleotide site-directed mutagenesis, no formation of stable hexamer, inactive mutant Solanum lycopersicum
3.4.11.1 E429D oligonucleotide site-directed mutagenesis, formation of a stable hexamer corresponding to the wild-type enzyme, activity with dipeptide substrates is similar or slightly reduced compared to the wild-type enzyme Solanum lycopersicum
3.4.11.1 E429G oligonucleotide site-directed mutagenesis, no formation of stable hexamer, inactive mutant Solanum lycopersicum
3.4.11.1 E429Q oligonucleotide site-directed mutagenesis, no formation of stable hexamer, inactive mutant Solanum lycopersicum
3.4.11.1 E429S oligonucleotide site-directed mutagenesis, formation of a stable hexamer corresponding to the wild-type enzyme Solanum lycopersicum
3.4.11.1 E429V oligonucleotide site-directed mutagenesis, formation of a stable hexamer corresponding to the wild-type enzyme, reduced catalytic activity with dipeptide substrates compared to the wild-type Solanum lycopersicum
3.4.11.1 E429W oligonucleotide site-directed mutagenesis, formation of a stable hexamer corresponding to the wild-type enzyme, nearly catalytically inactive with dipeptide substrates Solanum lycopersicum
3.4.11.1 K354E oligonucleotide site-directed mutagenesis, formation of a more complex structure compared to the wild-type enzyme Solanum lycopersicum
3.4.11.1 K354G oligonucleotide site-directed mutagenesis, formation of a more complex structure compared to the wild-type enzyme Solanum lycopersicum
3.4.11.1 K354M oligonucleotide site-directed mutagenesis, formation of a more complex structure compared to the wild-type enzyme Solanum lycopersicum
3.4.11.1 K354N oligonucleotide site-directed mutagenesis, formation of a more complex structure compared to the wild-type enzyme Solanum lycopersicum
3.4.11.1 K354R oligonucleotide site-directed mutagenesis, formation of a stable hexamer corresponding to the wild-type enzyme Solanum lycopersicum
3.4.11.1 K354T oligonucleotide site-directed mutagenesis, formation of a more complex structure compared to the wild-type enzyme Solanum lycopersicum
3.4.11.1 K354W oligonucleotide site-directed mutagenesis, formation of a more complex structure compared to the wild-type enzyme Solanum lycopersicum
3.4.11.1 R431A oligonucleotide site-directed mutagenesis, formation of a stable hexamer corresponding to the wild-type enzyme, nearly catalytically inactive with dipeptide substrates Solanum lycopersicum
3.4.11.1 R431G oligonucleotide site-directed mutagenesis, no formation of stable hexamer, inactive mutant Solanum lycopersicum
3.4.11.1 R431K oligonucleotide site-directed mutagenesis, reduced catalytic activity with dipeptide substrates compared to the wild-type Solanum lycopersicum
3.4.11.1 R431Q oligonucleotide site-directed mutagenesis, formation of a stable hexamer corresponding to the wild-type enzyme Solanum lycopersicum
3.4.11.1 R431V oligonucleotide site-directed mutagenesis, formation of a stable hexamer corresponding to the wild-type enzyme Solanum lycopersicum
3.4.11.1 R431W oligonucleotide site-directed mutagenesis, formation of a stable hexamer corresponding to the wild-type enzyme Solanum lycopersicum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.4.11.1 1.8
-
 L-Leu-Gly wild-type, pH 9.2, 37°C Solanum lycopersicum
3.4.11.1 2
-
 L-Leu-Gly mutant E429D, pH 9.2, 37°C Solanum lycopersicum
3.4.11.1 5.1
-
 L-Leu-Gly mutant R431K, pH 9.2, 37°C Solanum lycopersicum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.4.11.1 Mn2+
-
 Solanum lycopersicum
3.4.11.1 Zn2+ putative binding residues are Asp347 and Glu429 Solanum lycopersicum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.4.11.1 55000
-
 6 * 55000, wild-type Solanum lycopersicum

Organism

EC Number Organism UniProt Comment Textmining
3.4.11.1 Solanum lycopersicum
-
 wound-inducible enzyme
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.11.1 recombinant wild-type and mutant enzymes from Escherichia coli Solanum lycopersicum

Reaction

EC Number Reaction Comment Organism Reaction ID
3.4.11.1 release of an N-terminal amino acid, Xaa-/-Yaa-, in which Xaa is preferably Leu, but may be other amino acids including Pro although not Arg or Lys, and Yaa may be Pro. Amino acid amides and methyl esters are also readily hydrolysed, but rates on arylamides are exceedingly low catalytic residues are Arg431 and Lys354 Solanum lycopersicum
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.4.11.1 9.8
-
 purified recombinant wild-type enzyme, substrate L-Leu-L-Asp Solanum lycopersicum
3.4.11.1 45.7
-
 purified recombinant wild-type enzyme, substrate L-Tyr-L-Leu Solanum lycopersicum
3.4.11.1 112.3
-
 purified recombinant wild-type enzyme, substrate L-Leu-L-Arg Solanum lycopersicum
3.4.11.1 131.3
-
 purified recombinant wild-type enzyme, substrate L-Phe-L-Leu Solanum lycopersicum
3.4.11.1 141.6
-
 purified recombinant wild-type enzyme, substrate L-Leu-L-Asn Solanum lycopersicum
3.4.11.1 150.4
-
 purified recombinant wild-type enzyme, substrate L-Leu-L-Ser Solanum lycopersicum
3.4.11.1 150.9
-
 purified recombinant wild-type enzyme, substrate L-His-L-Leu Solanum lycopersicum
3.4.11.1 153.4
-
 purified recombinant wild-type enzyme, substrate L-Pro-L-Leu Solanum lycopersicum
3.4.11.1 160
-
 purified recombinant wild-type enzyme, substrate L-Leu-L-Gly Solanum lycopersicum
3.4.11.1 200.8
-
 purified recombinant wild-type enzyme, substrate L-Leu-L-Tyr Solanum lycopersicum
3.4.11.1 214.3
-
 purified recombinant wild-type enzyme, substrate L-Trp-L-Leu Solanum lycopersicum
3.4.11.1 261.3
-
 purified recombinant wild-type enzyme, substrate L-Thr-L-Leu Solanum lycopersicum
3.4.11.1 281.6
-
 purified recombinant wild-type enzyme, substrate L-Val-L-Leu Solanum lycopersicum
3.4.11.1 320.6
-
 purified recombinant wild-type enzyme, substrate L-Ala-L-Leu Solanum lycopersicum
3.4.11.1 325.4
-
 purified recombinant wild-type enzyme, substrate L-Leu-L-Met Solanum lycopersicum
3.4.11.1 345.3
-
 purified recombinant wild-type enzyme, substrate L-Met-L-Leu Solanum lycopersicum
3.4.11.1 419.3
-
 purified recombinant wild-type enzyme, substrate L-Leu-L-Leu Solanum lycopersicum
3.4.11.1 440.9
-
 purified recombinant wild-type enzyme, substrate L-Leu-L-Phe Solanum lycopersicum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.11.1 L-Ala-L-Leu + H2O
-
 Solanum lycopersicum L-Ala + L-Leu
-
 ?
3.4.11.1 L-His-L-Leu + H2O
-
 Solanum lycopersicum L-His + L-Leu
-
 ?
3.4.11.1 L-Leu-Gly + H2O
-
 Solanum lycopersicum L-Leu + Gly
-
 ?
3.4.11.1 L-Leu-L-Arg + H2O
-
 Solanum lycopersicum L-Leu + L-Arg
-
 ?
3.4.11.1 L-Leu-L-Asn + H2O
-
 Solanum lycopersicum L-Leu + L-Asn
-
 ?
3.4.11.1 L-Leu-L-Asp + H2O low activity Solanum lycopersicum L-Leu + L-Asp
-
 ?
3.4.11.1 L-Leu-L-Leu + H2O
-
 Solanum lycopersicum L-Leu + L-Leu
-
 ?
3.4.11.1 L-Leu-L-Met + H2O
-
 Solanum lycopersicum L-Leu + L-Met
-
 ?
3.4.11.1 L-Leu-L-Phe + H2O
-
 Solanum lycopersicum L-Leu + L-Phe
-
 ?
3.4.11.1 L-Leu-L-Ser + H2O
-
 Solanum lycopersicum L-Leu + L-Ser
-
 ?
3.4.11.1 L-Leu-L-Tyr + H2O
-
 Solanum lycopersicum L-Leu + L-Tyr
-
 ?
3.4.11.1 L-Met-L-Leu + H2O
-
 Solanum lycopersicum L-Met + L-Leu
-
 ?
3.4.11.1 L-Phe-L-Leu + H2O
-
 Solanum lycopersicum L-Phe + L-Leu
-
 ?
3.4.11.1 L-Pro-L-Leu + H2O
-
 Solanum lycopersicum L-Pro + L-Leu
-
 ?
3.4.11.1 L-Thr-L-Leu + H2O
-
 Solanum lycopersicum L-Thr + L-Leu
-
 ?
3.4.11.1 L-Trp-L-Leu + H2O
-
 Solanum lycopersicum L-Trp + L-Leu
-
 ?
3.4.11.1 L-Tyr-L-Leu + H2O
-
 Solanum lycopersicum L-Tyr + L-Leu
-
 ?
3.4.11.1 L-Val-L-Leu + H2O
-
 Solanum lycopersicum L-Val + L-Leu
-
 ?
3.4.11.1 Leu-4-nitroanilide + H2O
-
 Solanum lycopersicum Leu + 4-nitroaniline
-
 ?
3.4.11.1 additional information substrate specificty of wild-type and mutant enzymes, overview Solanum lycopersicum ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.4.11.1 hexamer 6 * 55000, wild-type Solanum lycopersicum
3.4.11.1 More quarternary structure of recombinant wild-type and mutant enzymes Solanum lycopersicum

Synonyms

EC Number Synonyms Comment Organism
3.4.11.1 LAP-A
-
 Solanum lycopersicum
3.4.11.1 leucine aminopeptidase
-
 Solanum lycopersicum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.4.11.1 37
-
 assay at Solanum lycopersicum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.4.11.1 30
-
 L-Leu-Gly mutant R431K, pH 9.2, 37°C Solanum lycopersicum
3.4.11.1 108
-
 L-Leu-Gly mutant E429D, pH 9.2, 37°C Solanum lycopersicum
3.4.11.1 120
-
 L-Leu-Gly wild-type, pH 9.2, 37°C Solanum lycopersicum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.4.11.1 9.2
-
 assay at Solanum lycopersicum