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Literature summary for 3.4.24.56 extracted from

  • Smith-Carpenter, J.E.; Alper, B.J.
    Functional requirement for human pitrilysin metallopeptidase 1 arginine 183, mutated in amyloidogenic neuropathy (2018), Protein Sci., 27, 861-873 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.24.56

Cloned(Commentary)

Cloned (Comment) Organism
expressed in and isolated from Escherichia coli as C-terminal polyhistidine tagged fusion partners lacking N-terminal mitochondrial targeting sequence Homo sapiens

Protein Variants

Protein Variants Comment Organism
E107Q catalytically inactive mutant Homo sapiens
R183a about 35% of the activity compared to wild-type enzyme with the substrate Homo sapiens
R183D less than 5% of the activity compared to wild-type enzyme with the substrate (7-methoxycoumarin-4-yl)acetyl-KLVFFAEDK(Dnp)-OH Homo sapiens
R183E about less than 5% of the activity compared to wild-type enzyme with the substrate (7-methoxycoumarin-4-yl)acetyl-KLVFFAEDK(Dnp)-OH Homo sapiens
R183K about 30% of the activity compared to wild-type enzyme with the substrate (7-methoxycoumarin-4-yl)acetyl-KLVFFAEDK(Dnp)-OH Homo sapiens
R183N about 10% of the activity compared to wild-type enzyme with the substrate (7-methoxycoumarin-4-yl)acetyl-KLVFFAEDK(Dnp)-OH Homo sapiens
R183Q mutant Pitrm1 R183Q is implicated in inherited amyloidogenic neuropathy. Recombinant R183Q mutant is less active than the recombinant wild-type enzyme against recombinant amyloid beta-peptide (Abeta1-40). R183Q mutant enzyme exhibits significantly decreased rate of fluorogenic peptide hydrolysis ((7-methoxycoumarin-4-yl)acetyl-KLVFFAEDK(Dnp)-OH), yet retains similar binding affinity by comparison with the wild-type enzyme. Residue R183 is positioned within an N-terminal strand-loop-strand motif that is essential for enzyme function. A requirement for charged residues within 4.5 A of residue R183 is demonstrated. The R183Q mutant enzyme exhibits increased sensitivity to heat inactivation Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0047
-
 7-methoxycoumarin-4-yl-acetyl-RPPGFSAFK-2,4-dinitrophenyl pH 7.0, 37°C, wild-type enzyme Homo sapiens
0.0049
-
 7-methoxycoumarin-4-yl-acetyl-RPPGFSAFK-2,4-dinitrophenyl pH 7.0, 37°C, mutant enzyme R183Q Homo sapiens
0.0056
-
 (7-methoxycoumarin-4-yl)acetyl-KLVFFAEDK(Dnp)-OH pH 7.0, 37°C, mutant enzyme R183Q Homo sapiens
0.0076
-
 (7-methoxycoumarin-4-yl)acetyl-KLVFFAEDK(Dnp)-OH pH 7.0, 37°C, wild-type enzyme Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens Q5JRX3
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(7-methoxycoumarin-4-yl)acetyl-KLVFFAEDK(Dnp)-OH + H2O fluorogenic substrate derived from the reported Abeta1-40 core peptide cleavage sequence. The R183Q mutant enzyme exhibits significantly decreased rate of fluorogenic peptide hydrolysis, yet retains similar binding affinity by comparison with the wild-type enzyme Homo sapiens ?
-
 ?
7-methoxycoumarin-4-yl-acetyl-RPPGFSAFK-2,4-dinitrophenyl + H2O
-
 Homo sapiens ?
-
 ?
amyloid beta-peptide (Abeta1-40) + H2O recombinant R183Q mutant enzyme is less active than the recombinant wild-type enzyme against recombinant amyloid beta-peptide (Abeta1-40) Homo sapiens ?
-
 ?

Synonyms

Synonyms Comment Organism
pitrilysin metallopeptidase 1
-
 Homo sapiens
Pitrm1
-
 Homo sapiens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
42
-
 3 min, 50% inactivation of mutant enzyme R183Q Homo sapiens
49
-
 3 min, 50% inactivation of wild-type enzyme Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.2
-
 7-methoxycoumarin-4-yl-acetyl-RPPGFSAFK-2,4-dinitrophenyl pH 7.0, 37°C, mutant enzyme R183Q Homo sapiens
1.2
-
 (7-methoxycoumarin-4-yl)acetyl-KLVFFAEDK(Dnp)-OH pH 7.0, 37°C, mutant enzyme R183Q Homo sapiens
6.5
-
 7-methoxycoumarin-4-yl-acetyl-RPPGFSAFK-2,4-dinitrophenyl pH 7.0, 37°C, wild-type enzyme Homo sapiens
7.5
-
 (7-methoxycoumarin-4-yl)acetyl-KLVFFAEDK(Dnp)-OH pH 7.0, 37°C, wild-type enzyme Homo sapiens