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Literature summary for 3.4.11.1 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
Search for more literature for 3.4.11.1

Cloned(Commentary)

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

Protein Variants

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

KM Value [mM]

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

Metals/Ions

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

Molecular Weight [Da]

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

Organism

Organism UniProt Comment Textmining
Solanum lycopersicum
-
 wound-inducible enzyme
-

Purification (Commentary)

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

Reaction

Reaction Comment Organism Reaction ID
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]

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

Synonyms Comment Organism
LAP-A
-
 Solanum lycopersicum
leucine aminopeptidase
-
 Solanum lycopersicum

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

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