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Information on EC 3.4.24.33 - peptidyl-Asp metalloendopeptidase
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EC Tree 3 Hydrolases
3.4 Acting on peptide bonds (peptidases)
3.4.24 Metalloendopeptidases
3.4.24.33 peptidyl-Asp metalloendopeptidase
IUBMB Comments
A metalloenzyme isolated from Pseudomonas fragi. Useful in protein sequencing applications because of its limited specificity. In peptidase family M72.
The enzyme appears in viruses and cellular organisms
showSynonyms
endoproteinase asp-n, asp-n, asp-n endoproteinase, endoproteinase aspn, endoprotease asp-n, peptidyl-asp metalloendopeptidase, more
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
AspN
Endoproteinase Asp-N
Peptidyl-Asp metalloproteinase
-
-
-
-
Proteinase, peptidyl-Asp metallo-
-
-
-
-
Endoproteinase Asp-N
-
-
-
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Cleavage of Xaa-/-Asp, Xaa-/-Glu and Xaa-/-cysteic acid bonds
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-
-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of peptide bond
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CAS REGISTRY NUMBER
COMMENTARY
55576-49-3
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
alpha-tubulin + H2O
?
-
Substrates: specific for preaspartate cleavage
Products: -
Products: -
?
antithrombin + H2O
?
-
Substrates: the Asp342 localized in helix I is the AspN cleavage site, found in both heating-induced and citrullination-induced polymers of antithrombin
Products: -
Products: -
?
Apolipoprotein A-I + H2O
Peptides
-
Substrates: 2 CNBr-fragments, cleavage at 12 Asp-residues and 5 out of 18 Glu-residues, cleaves N-terminal to Glu as well as to Asp and cysteic acid
Products: amino acid sequences
Products: amino acid sequences
?
Erythrocyte carbonic anhydrase I + H2O
?
-
Substrates: i.e. EC 4.2.1.1, cleavage at 5 Asp- and 1 Glu-residues
Products: -
Products: -
?
Human erythrocyte D-aspartyl-L-isoaspartyl methyltransferase isozyme I + H2O
?
-
Substrates: i.e. EC 2.1.1.77, cleavage sites: N-terminal side of Asp and 5 out of 9 Glu-residues
Products: -
Products: -
?
recombinant histone H1.3 + H2O
C-terminal fragment N.1 of recombinant histone H1.3 + ?
-
Substrates: -
Products: -
Products: -
?
casein + H2O
?
-
Substrates: specific for preaspartate cleavage
Products: -
Products: -
?
Glucagon + H2O
?
-
Substrates: specific for preaspartate cleavage
Products: -
Products: -
?
Insulin B-chain + H2O
?
-
Substrates: specific for preaspartate cleavage
Products: -
Products: -
?
Insulin B-chain + H2O
?
-
Substrates: oxidized with performic acid
Products: -
Products: -
?
Insulin B-chain + H2O
?
-
Substrates: cleavage sites: Leu6-Cys7, Leu15-Tyr16, Val18-Cys19, Phe24-Phe25
Products: -
Products: -
?
Insulin B-chain + H2O
?
-
Substrates: also cleaves bonds with cysteic acid in P1' derived from cysteine residues by oxidation with performic acid and at N-terminal side of some glutamyl residues
Products: -
Products: -
?
Myoglobin + H2O
?
-
Substrates: AspN and 0.5 microg/microl myoglobin at an enzyme to substrate ratio of 1:70 w/w
Products: -
Products: -
?
Myoglobin + H2O
?
-
Substrates: AspN and 0.5 microg/microl myoglobin at an enzyme to substrate ratio of 1:70 w/w
Products: -
Products: -
?
Pancreatic ribonuclease + H2O
Peptides
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Substrates: oxidized with performic acid, cleavage sites: -Xaa-Asp- and -Xaa-Cys- (not Cys40, Cys84 or Glu-residues)
Products: i.e. peptide(1-13), peptide(14-25), peptide(26-37), peptide(38-52), peptide(53-57), peptide(53-64), peptide(65-71), peptide(72-82), peptide(83-94), peptide(110-120), peptide(121-124)
Products: i.e. peptide(1-13), peptide(14-25), peptide(26-37), peptide(38-52), peptide(53-57), peptide(53-64), peptide(65-71), peptide(72-82), peptide(83-94), peptide(110-120), peptide(121-124)
?
Pancreatic ribonuclease + H2O
Peptides
-
Substrates: oxidized with performic acid, cleavage sites: -Xaa-Asp- and -Xaa-Cys- (not Cys40, Cys84 or Glu-residues)
Products: i.e. peptide(1-13), peptide(14-25), peptide(26-37), peptide(38-52), peptide(53-57), peptide(53-64), peptide(65-71), peptide(72-82), peptide(83-94), peptide(110-120), peptide(121-124)
Products: i.e. peptide(1-13), peptide(14-25), peptide(26-37), peptide(38-52), peptide(53-57), peptide(53-64), peptide(65-71), peptide(72-82), peptide(83-94), peptide(110-120), peptide(121-124)
?
Sperm whale myoglobin + H2O
?
-
Substrates: in the presence of 2 M urea cleavage of 4 out of 6 Asp-residues
Products: -
Products: -
?
Sperm whale myoglobin + H2O
?
-
Substrates: in the presence of 2 M urea cleavage of 4 out of 6 Asp-residues
Products: -
Products: -
?
additional information
?
-
-
Substrates: the wild-type protease has no well-delineated specificity, some preference for N-terminal side of hydrophilic residues, e.g. aminoethylcysteine, Ser, Thr, Gln and Gly
Products: -
Products: -
?
additional information
?
-
-
Substrates: high substrate specificity of AspN, that ensures that all of the non-N-terminal peptides having aspartic acid or glutamic acid at their N-termini can be converted. An artificially targeted N-blocked protein is digested with AspN, method overview. The proposed method is applicable to proteins, whether N blocked or N free
Products: -
Products: -
?
additional information
?
-
-
Substrates: endoprotease Asp-N selectively cleaves aspartyl peptides but not the isoaspartyl counterparts
Products: -
Products: -
?
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
data obtained from inhibition experiments support metalloproteinase character
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Alpha-macroglobulin
-
inhibition at a molar ratio of inhibitor to protease of about 18 to 1
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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DISEASE
TITLE OF PUBLICATION
LINK TO PUBMED
Cataract
An atypical form of alphaB-crystallin is present in high concentration in some human cataractous lenses. Identification and characterization of aberrant N- and C-terminal processing.
Diabetes Mellitus, Type 2
Quantitative analytical method for determining the levels of gastric inhibitory polypeptides GIP1-42 and GIP3-42 in human plasma using LC-MS/MS/MS.
Measles
Isolation of the measles virus hemagglutinin protein in a soluble form by protease digestion.
Neoplasms
Biological and clinical relevance of the urokinase-type plasminogen activator (uPA) in breast cancer.
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
obtained by growth of Pseudomonas fragi ATCC 4973 on elastin as sole C-source
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-
brenda
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Highest Expressing Human Cell Lines
Cell Line LinksPlease wait a moment until the data is sorted. This message will disappear when the data is sorted.
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). ASPN_STRMK
Stenotrophomonas maltophilia (strain K279a)
417
0
44141
Swiss-Prot
-
ASPN_STRM5
Stenotrophomonas maltophilia (strain R551-3)
417
0
44067
Swiss-Prot
-
ASPN_XANCP
Xanthomonas campestris pv. campestris (strain ATCC 33913 / DSM 3586 / NCPPB 528 / LMG 568 / P 25)
417
0
44137
Swiss-Prot
-
MEP72_PSEAE
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
599
0
64960
Swiss-Prot
-
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
27000
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
monomer
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
derepressed mutant of Pseudomonas fragi ATCC 4973 produces 40 times higher proteinase levels
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
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ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, lyophilized in the presence of Tris-HCl buffer, pH 7.5, at least 2 years, 4 mg lyophilizate in 100 ml H2O, 2 weeks
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
proteolytic digestion of alpha-parvalbumin using endoproteinase Asp-N for subsequent examination by MALDI-TOF
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
analysis
-
analysis of isoaspartic acid by selective proteolysis with Asp-N and electron transfer dissociation mass spectrometry, overview. IsoAsp formation and repair is central to the survival and germination of plant seeds. Also once administered into patients and thus exposed to physiological conditions of pH 7 and 37 °C, protein pharmaceuticals, particularly those with long circulation time, may generate significant amount of isoAsp
molecular biology
-
many eukaryotic proteins are blocked at the alpha-amino group of their N-terminal with various modifications, thereby making it difficult to determine their N-terminal sequence by protein sequencer, development of a method for selectively isolating the blocked N-terminal peptide from the peptide mixture generated by endoproteinase AspN digestion of N-blocked protein by removal of all peptides other than the N-terminal one (non-N-terminal peptides) through their carbonyl group introduced by a chemical transamination reaction
additional information
additional information
-
AspN is shown to be an alternative protease for in-capillary digestion (during capillary electrophoresis), AspN behaves very similarly to trypsin
additional information
-
AspN is shown to be an alternative protease for in-capillary digestion (during capillary electrophoresis), AspN behaves very similarly to trypsin
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Ingrosso, D.; Fowler, A.V.; Bleibaum, J.; Clarke, S.
Specificity of endoproteinase Asp-N (Pseudomonas fragi): cleavage at glutamyl residues in two proteins
Biochem. Biophys. Res. Commun.
162
1528-1534
1989
Pseudomonas fragi
brenda
Noreau, J.; Drapeau, G.R.
Isolation and properties of the protease from the wild-type and mutant strains of Pseudomonas fragi
J. Bacteriol.
140
911-916
1979
Pseudomonas fragi
brenda
Drapeau, G.R.
Substrate specificity of a proteolytic enzyme isolated from a mutant of Pseudomonas fragi
J. Biol. Chem.
255
839-840
1980
Pseudomonas fragi, Pseudomonas fragi Me1
brenda
Gagmann, M.L.; Geuss, U.; Fischer, S.; Kresse, G.B.
Peptidyl-Asp metalloendopeptidase
Methods Enzymol.
248
782-787
1995
Pseudomonas fragi
brenda
Tetaz, T.; Morrison, J.R.; Andreou, J.; Fidge, N.H.
Relaxed specificity of endoproteinase Asp-N: this enzyme cleaves at peptide bonds N-terminal to glutamate as well as aspartate and cysteic acid residues
Biochem. Int.
22
561-566
1990
Pseudomonas fragi
brenda
Hagmann, M.
Peptidyl-Asp metalloendopeptidase
Handbook of Proteolytic Enzymes(Barrett,A. J. ,Rawlings,N. D. ,Woessner,J. F. ,Eds. )Academic Press
1
1037-1039
2004
Pseudomonas fragi
-
brenda
Nesbitt, C.A.; Yeung, K.K.
In-capillary enrichment, proteolysis and separation using capillary electrophoresis with discontinuous buffers: application on proteins with moderately acidic and basic isoelectric points
Analyst
134
65-71
2009
Pseudomonas fragi, Pseudomonas fragi mutant
brenda
Permyakov, S.E.; Karnoup, A.S.; Bakunts, A.G.; Permyakov, E.A.
Sequence microheterogeneity of parvalbumin pI 5.0 of pike: a mass spectrometric study
Biochim. Biophys. Acta
1794
129-136
2009
Pseudomonas fragi
brenda
Ordonez, A.; Martinez-Martinez, I.; Corrales, F.J.; Miqueo, C.; Minano, A.; Vicente, V.; Corral, J.
Effect of citrullination on the function and conformation of antithrombin
FEBS J.
276
6763-6772
2009
Pseudomonas fragi
brenda
Seo, J.H.; Lim, J.C.; Lee, D.Y.; Kim, K.S.; Piszczek, G.; Nam, H.W.; Kim, Y.S.; Ahn, T.; Yun, C.H.; Kim, K.; Chock, P.B.; Chae, H.Z.
Novel protective mechanism against irreversible hyperoxidation of peroxiredoxin: Nalpha-terminal acetylation of human peroxiredoxin II
J. Biol. Chem.
284
13455-13465
2009
Pseudomonas fragi
brenda
Soslau, G.; Prest, P.J.; Class, R.; Jost, M.; Mathews, L.
Inhibition of gamma-thrombin-induced human platelet aggregation by histone H1 subtypes and H1.3 fragments
Platelets
20
349-356
2009
Vibrio proteolyticus
brenda
Sonomura, K.; Kuyama, H.; Matsuo, E.; Tsunasawa, S.; Futaki, S.; Nishimura, O.
Selective isolation of N-blocked peptide by combining AspN digestion, transamination, and tosylhydrazine glass treatment
Anal. Biochem.
410
214-223
2011
Pseudomonas fragi
brenda
Ni, W.; Dai, S.; Karger, B.L.; Zhou, Z.S.
Analysis of isoaspartic acid by selective proteolysis with Asp-N and electron transfer dissociation mass spectrometry
Anal. Chem.
82
7485-7491
2010
Pseudomonas fragi
brenda
EXTERNAL LINKS (specific for EC-Number 3.4.24.33)
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