Cloned (Comment) | Organism |
---|---|
gene LNPEP, recombinant expression of His-tagged wild-type and mutant enzymes in Spodoptera frugiperda Sf9 cells and Trichoplusia ni Hi5 cells via the baculovirus transfection system | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme free or in complex with inhibitory antigenic peptide precursor analogue N2-(2- 8[(1-amino-3-phenylpropyl)(hydroxy)phosphoryl]methyl]-4-methylpentanoyl)-L-lysyl-L-histidyl-L-histidyl-L-alanyl-L-phenylalanyl-L-seryl-L-phenylalanyl-L-lysine, sitting drop vapor diffusion, mixing of 100 nl of 10 mg/ml protein in 150 mM NaCl and 10 mM HEPES, pH 7.4, with 100 nl of crystallization solution containing 10% w/v PEG 4000, 20% v/v glycerol, 53.4 mM bicine, 46.6 mM Trizma base, pH 8.5, and 0.02 M each of sodium L-glutamate, DL-alanine, glycine, DL-lysine, and DL-serine, method optimization, X-ray diffraction structure determination and analysis at 3.31-3.37 A resolution, molecular replacement using the highly homologous ERAP1 open structure (PDB ID 3QNF) as a search model, and modelling | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
A6909T | naturally occuing mutation, polymorphism A609T interacts with the hinge domain III of IRAP, similarly to SNP K528R in ERAP1, which has been repeatedly validated for disease association and effects on activity. The A609T mutation leads to an almost 2fold reduction in activity compared to wild-type | Homo sapiens |
I166M | naturally occuing mutation, the I166M polymorphism is located in domain I of the enzyme at the interface with domain II and makes contact with loop 534-538 that is adjacent to the S1 pocket of the enzyme. The mutant enzyme activity is slightly reduced compared to the wild-type | Homo sapiens |
additional information | mapping and functional characterization of IRAP disease-associated singel-nucleotide polymorphisms, SNPs. Two coding single-nucleotide polymorphisms in IRAP have been associated with predisposition to the autoimmune diseases psoriasis and ankylosing spondylitis, namely rs2303138 coding for the A609T change and rs61752351 coding for the I166M change | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
N2-(2- 8[(1-amino-3-phenylpropyl)(hydroxy)phosphoryl]methyl]-4-methylpentanoyl)-L-lysyl-L-histidyl-L-histidyl-L-alanyl-L-phenylalanyl-L-seryl-L-phenylalanyl-L-lysine | i.e. DG025, an antigenic peptide precursor analogue, enzyme binding structure determination and analysis. The canonical orientation of the two N-terminal residues of DG025 is defined by the transition state nature of the ligand and the hydrophobic and aromatic interactions of the hPhe residue in the S1 specificity pocket of the enzyme (and in particular with Phe544). The remaining of the peptide extends toward the base of the cavity, where it is sandwiched between domains II and IV | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q9UIQ6 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Trichoplusia ni Hi5 cells by dialysis and nickel affinity chromatography | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
dendritic cell | - |
Homo sapiens | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-leucine-7-amido-4-methylcoumarin + H2O | - |
Homo sapiens | L-leucine + 7-amino-4-methylcoumarin | - |
? | |
LGGGGGL + H2O | hydrolysis of poly-glycine peptides, overview | Homo sapiens | L-Leu + GGGGGL | - |
? |
Synonyms | Comment | Organism |
---|---|---|
insulin-regulated aminopeptidase | - |
Homo sapiens |
IRAP | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Homo sapiens |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.0000077 | - |
pH 7.0, 37°C, recombinant wild-type enzyme | Homo sapiens | N2-(2- 8[(1-amino-3-phenylpropyl)(hydroxy)phosphoryl]methyl]-4-methylpentanoyl)-L-lysyl-L-histidyl-L-histidyl-L-alanyl-L-phenylalanyl-L-seryl-L-phenylalanyl-L-lysine |
General Information | Comment | Organism |
---|---|---|
physiological function | aminopeptidases generate antigenic peptides influence immunodominance and adaptive cytotoxic immune responses. These enzymes efficiently process a vast number of different long peptide substrates. Insulin-regulated aminopeptidase prepares antigenic epitopes for crosspresentation in dendritic cells | Homo sapiens |