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Literature summary for 3.4.24.56 extracted from

  • Noinaj, N.; Bhasin, S.K.; Song, E.S.; Scoggin, K.E.; Juliano, M.A.; Juliano, L.; Hersh, L.B.; Rodgers, D.W.
    Identification of the allosteric regulatory site of insulysin (2011), PLoS ONE, 6, e20864.
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
ATP 40fold activation for wild-type Rattus norvegicus
bradykinin 4fold activation for wild-type Rattus norvegicus

Cloned(Commentary)

Cloned (Comment) Organism
expressed as hexahistidine fusion proteins in SF9 insect cell Rattus norvegicus

Protein Variants

Protein Variants Comment Organism
E111F crystal structure of a catalytically compromised E111F mutant of IDE has electron density for peptide ligands bound at the active site in domain 1 and a distal site in domain 2 Rattus norvegicus
I374S mutation in the distal site eliminates allosterism, Vmax decreased approximately 10fold compared to wild-type, Km (Abz-GGFLRKHGQ-EDDnp) roughly similar to wild-type, no heterotropic activation with bradykinin, no significant activation through ATP Rattus norvegicus
V360S mutation in the distal site eliminates allosterism, Vmax decreased approximately 10fold compared to wild-type, Km (Abz-GGFLRKHGQ-EDDnp) roughly similar to wild-type, no heterotropic activation with bradykinin, ATP activation is reduced to 8fold Rattus norvegicus
Y609F mutation in the distal site eliminates allosterism, Vmax decreased approximately 10fold compared to wild-type, Km (Abz-GGFLRKHGQ-EDDnp) roughly similar to wild-type, no heterotropic activation with bradykinin, no significant activation through ATP Rattus norvegicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0068
-
Abz-GGFLRKHGQ-EDDnp pH 7.4, temperature not specified in the publication, mutant T609F, Vmax: 2.6 nmol/min/mg Rattus norvegicus
0.0085
-
Abz-GGFLRKHGQ-EDDnp pH 7.4, temperature not specified in the publication, mutant I374S, Vmax: 3.45 nmol/min/mg Rattus norvegicus
0.00861
-
Abz-GGFLRKHGQ-EDDnp pH 7.4, temperature not specified in the publication, mutant V360S, Vmax: 2 nmol/min/mg Rattus norvegicus
0.01587
-
Abz-GGFLRKHGQ-EDDnp pH 7.4, temperature not specified in the publication, wild-type, Vmax: 35.5 nmol/min/mg Rattus norvegicus

Organism

Organism UniProt Comment Textmining
Rattus norvegicus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
using Ni-NTA chromatography Rattus norvegicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Abz-GGFLRKHGQ-EDDnp + H2O
-
Rattus norvegicus Abz-GGFLR + KHGQ-EDDnp
-
?

Subunits

Subunits Comment Organism
dimer
-
Rattus norvegicus

Synonyms

Synonyms Comment Organism
IDE
-
Rattus norvegicus
Insulysin
-
Rattus norvegicus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
assay at Rattus norvegicus