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Literature summary for 3.5.1.60 extracted from
- Mass spectrometric characterization of human N-acylethanolamine-hydrolyzing acid midase (2012), J. Proteome Res., 11, 972-981.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| additional information | the enzyme is proteolytically activated by an autocatalytic step under acidic conditions where the polypeptide is cleaved into two chains | Homo sapiens |
Application
| Application | Comment | Organism |
|---|---|---|
| pharmacology | potential of blocking N-acylethanolamines like palmitoylethanolamide or N-arachidonlyethanolamine (anandamide) from enzymatic degradation via enzyme inhibition as a strategy for pain treatment | Homo sapiens |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| recombinant stable expression of the His6-tagged zymogen in HEK-293 cells, ammonium chloride treatment stimulates secretion of the lysosomal proteins from the HEK293 cells | Homo sapiens |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| additional information | selective alkylation of the enzyme's cysteine residues results in almost complete loss of activity | Homo sapiens |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| additional information | selective alkylation of the enzyme's cysteine residues results in almost complete loss of activity | Homo sapiens |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Michaelis-Menten kinetics | Homo sapiens | |
| 0.0062 | - |
N-(4-methylcoumarin)palmitamide | pH 4.5, 37°C | Homo sapiens |  |
| 0.021 | - |
palmitoylethanolamide | pH 4.5, 37°C | Homo sapiens | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| lysosome | the enzyme is believed to be transported by the mannose-6-phosphate pathway to the acidic late endosomes and/or lysosomes | Homo sapiens | 5764 | - |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 10972 | - |
1 * 10972, deglycosylated alpha-subunit, + 1 * 29659, deglycosylated beta-subunit, mass spectrometry, 1 * 14600 + 1 * 33000, alpha/beta heterodimer, SDS-PAGE | Homo sapiens |
| 14600 | - |
1 * 10972, deglycosylated alpha-subunit, + 1 * 29659, deglycosylated beta-subunit, mass spectrometry, 1 * 14600 + 1 * 33000, alpha/beta heterodimer, SDS-PAGE | Homo sapiens |
| 29659 | - |
1 * 10972, deglycosylated alpha-subunit, + 1 * 29659, deglycosylated beta-subunit, mass spectrometry, 1 * 14600 + 1 * 33000, alpha/beta heterodimer, SDS-PAGE | Homo sapiens |
| 33000 | - |
1 * 10972, deglycosylated alpha-subunit, + 1 * 29659, deglycosylated beta-subunit, mass spectrometry, 1 * 14600 + 1 * 33000, alpha/beta heterodimer, SDS-PAGE | Homo sapiens |
| 40593 | - |
1 * 40593, deglycosylated zymogen, mass spectrometry | Homo sapiens |
| 45000 | - |
gel filtration | Homo sapiens |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Homo sapiens | Q02083 | - |
- |
Posttranslational Modification
| Posttranslational Modification | Comment | Organism |
|---|---|---|
| glycoprotein | 4 glycosylation sites Asn37, Asn107, Asn309, and Asn333, N-linked oligosaccharides of approximately 1500 Da, all cleavable by peptide-N-glycosidase F | Homo sapiens |
| proteolytic modification | the glycoprotein undergoes removal of an N-terminal signal peptide after biosynthesis. Autocatalytic acid cleavage of the zymogen into alpha- and beta-subunits (14.6 and 33.3 kDa) activates the enzyme. Cys126 is essential for the proteolytic cleavage of the pro-enzyme | Homo sapiens |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His6-tagged zymogen from HEK-293 cells by ammonium sulfate fractionation, dialysis, metal affinity chromatography, and again dialysis followed by ultrafiltration | Homo sapiens |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| N-(4-methylcoumarin)palmitamide + H2O | - |
Homo sapiens | palmitic acid + 7-amino-4-methylcoumarin | - |
? | |
| palmitoylethanolamide + H2O | - |
Homo sapiens | palmitic acid + ethanolamine | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| heterodimer | 1 * 10972, deglycosylated alpha-subunit, + 1 * 29659, deglycosylated beta-subunit, mass spectrometry, 1 * 14600 + 1 * 33000, alpha/beta heterodimer, SDS-PAGE | Homo sapiens |
| monomer | 1 * 40593, deglycosylated zymogen, mass spectrometry | Homo sapiens |
| More | tryptic digestion and MALDI-TOF mass spectrometry fingerprinting gives evidence for the lack of a disulfide bond between subunits | Homo sapiens |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| N-acylethanolamine-hydrolyzing acid amidase | - |
Homo sapiens |
| NAAA | - |
Homo sapiens |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Homo sapiens |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 4.5 | - |
assay at | Homo sapiens |
General Information
| General Information | Comment | Organism |
|---|---|---|
| additional information | the catalytic triad, is formed by cysteine, aspartate and arginine | Homo sapiens |
| physiological function | N-acylethanolamine-hydrolyzing acid amidase is a lysosomal enzyme that primarily degrades palmitoylethanolamine, a lipid amide that inhibits inflammatory responses | Homo sapiens |
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