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Literature summary extracted from

  • West, J.; Zvonok, N.; Whitten, K.; Wood, J.; Makriyannis, A.
    Mass spectrometric characterization of human N-acylethanolamine-hydrolyzing acid midase (2012), J. Proteome Res., 11, 972-981.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.5.1.60 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

EC Number Application Comment Organism
3.5.1.60 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)

EC Number Cloned (Comment) Organism
3.5.1.60 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

EC Number Protein Variants Comment Organism
3.5.1.60 additional information selective alkylation of the enzyme's cysteine residues results in almost complete loss of activity Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.1.60 additional information selective alkylation of the enzyme's cysteine residues results in almost complete loss of activity Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.60 additional information
-
 additional information Michaelis-Menten kinetics Homo sapiens
3.5.1.60 0.0062
-
 N-(4-methylcoumarin)palmitamide pH 4.5, 37°C Homo sapiens
3.5.1.60 0.021
-
 palmitoylethanolamide pH 4.5, 37°C Homo sapiens

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.5.1.60 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]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.1.60 10972
-
 1 * 10972, deglycosylated alpha-subunit, + 1 * 29659, deglycosylated beta-subunit, mass spectrometry, 1 * 14600 + 1 * 33000, alpha/beta heterodimer, SDS-PAGE Homo sapiens
3.5.1.60 14600
-
 1 * 10972, deglycosylated alpha-subunit, + 1 * 29659, deglycosylated beta-subunit, mass spectrometry, 1 * 14600 + 1 * 33000, alpha/beta heterodimer, SDS-PAGE Homo sapiens
3.5.1.60 29659
-
 1 * 10972, deglycosylated alpha-subunit, + 1 * 29659, deglycosylated beta-subunit, mass spectrometry, 1 * 14600 + 1 * 33000, alpha/beta heterodimer, SDS-PAGE Homo sapiens
3.5.1.60 33000
-
 1 * 10972, deglycosylated alpha-subunit, + 1 * 29659, deglycosylated beta-subunit, mass spectrometry, 1 * 14600 + 1 * 33000, alpha/beta heterodimer, SDS-PAGE Homo sapiens
3.5.1.60 40593
-
 1 * 40593, deglycosylated zymogen, mass spectrometry Homo sapiens
3.5.1.60 45000
-
 gel filtration Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.60 Homo sapiens Q02083
-
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
3.5.1.60 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
3.5.1.60 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)

EC Number Purification (Comment) Organism
3.5.1.60 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)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.60 N-(4-methylcoumarin)palmitamide + H2O
-
 Homo sapiens palmitic acid + 7-amino-4-methylcoumarin
-
 ?
3.5.1.60 palmitoylethanolamide + H2O
-
 Homo sapiens palmitic acid + ethanolamine
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.1.60 heterodimer 1 * 10972, deglycosylated alpha-subunit, + 1 * 29659, deglycosylated beta-subunit, mass spectrometry, 1 * 14600 + 1 * 33000, alpha/beta heterodimer, SDS-PAGE Homo sapiens
3.5.1.60 monomer 1 * 40593, deglycosylated zymogen, mass spectrometry Homo sapiens
3.5.1.60 More tryptic digestion and MALDI-TOF mass spectrometry fingerprinting gives evidence for the lack of a disulfide bond between subunits Homo sapiens

Synonyms

EC Number Synonyms Comment Organism
3.5.1.60 N-acylethanolamine-hydrolyzing acid amidase
-
 Homo sapiens
3.5.1.60 NAAA
-
 Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.1.60 37
-
 assay at Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.1.60 4.5
-
 assay at Homo sapiens

General Information

EC Number General Information Comment Organism
3.5.1.60 additional information the catalytic triad, is formed by cysteine, aspartate and arginine Homo sapiens
3.5.1.60 physiological function N-acylethanolamine-hydrolyzing acid amidase is a lysosomal enzyme that primarily degrades palmitoylethanolamine, a lipid amide that inhibits inflammatory responses Homo sapiens