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Literature summary for 3.4.11.2 extracted from

  • Adang, M.J.
    Insect aminopeptidase N (2004), Handbook of Proteolytic Enzymes (Barrett, J. ; Rawlings, N. D. ; Woessner, J. F. , eds. ), 1, 296-299.
No PubMed abstract available

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli, which is more difficult for the full-length enzyme as for fragments, successfull expression of isozyme APN1 in Spodoptera frugiperda Sf9 cells using the baculovirus transfection system, but not of isozyme APN2, transient expression of isozyme APN1 on the cell surface in Sf21 cells under control of the inducible Drosophila HSP70 promoter Plutella xylostella

Protein Variants

Protein Variants Comment Organism
additional information the slamdance mutant, sda, has a modification of the APN structural gene, mutant flies exhibit altered neuronal excitability and are prone to seizures Drosophila melanogaster

Inhibitors

Inhibitors Comment Organism Structure
1,10-phenanthroline isozyme APN1 Bombyx mori
2,2'-dipyridyl isozyme APN1 Bombyx mori
actinonin isozyme APN4: IC50 is 0.148 mM, inhibits also isozyme APN1 Bombyx mori
amastatin isozyme APN4: IC50 is 0.0076 mM, inhibits also isozyme APN1 Bombyx mori
amastatin strong inhibition Tenebrio molitor
bestatin
-
Tenebrio molitor
probestin isozyme APN4: IC50 is 0.074 mM, inhibits also isozyme APN1 Bombyx mori

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane brush border Manduca sexta 16020
-
membrane brush border Plutella xylostella 16020
-

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ metallopeptidase Bombyx mori
Zn2+ the enzyme possesses a central Zn2+ coordination site H1EXXHX18E24 Heliothis virescens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
90000
-
-
Tenebrio molitor
110000
-
isozyme APN4 Bombyx mori
120000
-
isozyme APN1 Bombyx mori
120000
-
isozyme APN1 Epiphyas postvittana
120000 170000
-
Heliothis virescens
130000
-
recombinant isozyme APN1 from Sf9 cells Plutella xylostella

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Drosophila melanogaster the enzyme plays a role in behaviour and nervous system excitability ?
-
?
additional information Helicoverpa punctigera the isozymes have different roles in digestion ?
-
?

Organism

Organism UniProt Comment Textmining
Bombyx mori
-
-
-
Drosophila melanogaster
-
-
-
Epiphyas postvittana
-
-
-
Helicoverpa punctigera
-
isozymes APN1-4
-
Heliothis virescens
-
-
-
Manduca sexta
-
-
-
Plutella xylostella
-
-
-
Tenebrio molitor
-
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein
-
Tenebrio molitor
glycoprotein about 30% carbohydrate content, isozyme APN1 is rich in serine and threonine residues, potential O-glycosylation sites, near the C-terminus Heliothis virescens

Purification (Commentary)

Purification (Comment) Organism
by preparation of brush border membrane vesicles and wheat germ agglutinin affinity chromatography Heliothis virescens
by preparation of brush border membrane vesicles from whole larvae, anion exchange and hydrophobic interaction chromatography, or separation by affinity binding to Cry1Ac toxin Manduca sexta
partially by preparation of brush border membrane vesicles from whole larvae Plutella xylostella
separation by affinity binding of brush border membrane vesicle enzyme to Cry1Ac toxin Bombyx mori
to homogeneity Tenebrio molitor

Source Tissue

Source Tissue Comment Organism Textmining
central nervous system
-
Drosophila melanogaster
-
larva
-
Manduca sexta
-
larva
-
Plutella xylostella
-
larva expression of isozymes APN1 and APN2 during larval development in neostatestage and third larval stage and during the entire development, respectively Helicoverpa punctigera
-
midgut
-
Manduca sexta
-
midgut
-
Plutella xylostella
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Gly-4-nitroanilide + H2O low activity Bombyx mori Gly + 4-nitroaniline
-
?
L-Ala-4-nitroanilide + H2O
-
Bombyx mori L-Ala + 4-nitroaniline
-
?
L-Arg-4-nitroanilide + H2O
-
Bombyx mori L-Arg + 4-nitroaniline
-
?
L-Leu-4-nitroanilide + H2O
-
Bombyx mori L-Leu + 4-nitroaniline
-
?
L-Lys-4-nitroanilide + H2O
-
Bombyx mori L-Lys + 4-nitroaniline
-
?
L-Met-4-nitroanilide + H2O preferred substrate Bombyx mori L-Met + 4-nitroaniline
-
?
L-Pro-4-nitroanilide + H2O low activity Bombyx mori L-Pro + 4-nitroaniline
-
?
L-Val-4-nitroanilide + H2O low activity Bombyx mori L-Val + 4-nitroaniline
-
?
additional information the enzyme plays a role in behaviour and nervous system excitability Drosophila melanogaster ?
-
?
additional information the isozymes have different roles in digestion Helicoverpa punctigera ?
-
?
additional information broad substrate specificity for N-terminal amino acid residues, no activity with Asp-2-naphthylamide, the enzyme prefers tri- and tetrapeptides to di- and pentapeptides Tenebrio molitor ?
-
?
additional information isozyme APN1 shows preference for leucine amides, but also hydrolyzes Ile, Trp, Phe, Val, and His amides, no activity with Ser, Pro, Ala, Gly, and ARg derivatives Epiphyas postvittana ?
-
?
additional information the enzyme binds the Cry1Ac toxin from Bacillus thuringiensis Manduca sexta ?
-
?
additional information the enzyme prefers substrates with N-terminal nonpolar amino acids, isoyzme APN1 binds the Cry1Ac toxin from Bacillus thuringiensis via an N-acetylglucosamine moiety Bombyx mori ?
-
?

Subunits

Subunits Comment Organism
More the enzyme has a C-terminal anchor, and the N-terminal motif Trp-Arg-Leu-Pro Tenebrio molitor

Synonyms

Synonyms Comment Organism
aminopeptidase N
-
Drosophila melanogaster
aminopeptidase N
-
Bombyx mori
aminopeptidase N
-
Tenebrio molitor
aminopeptidase N
-
Manduca sexta
aminopeptidase N
-
Plutella xylostella
aminopeptidase N
-
Heliothis virescens
aminopeptidase N
-
Epiphyas postvittana
aminopeptidase N
-
Helicoverpa punctigera
APN
-
Drosophila melanogaster
APN
-
Tenebrio molitor
APN
-
Manduca sexta
APN
-
Helicoverpa punctigera
APN1
-
Bombyx mori
APN1
-
Plutella xylostella
APN1
-
Heliothis virescens
APN1
-
Epiphyas postvittana

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
-
Tenebrio molitor

pI Value

Organism Comment pI Value Maximum pI Value
Tenebrio molitor
-
-
7.4

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.0076
-
isozyme APN4: IC50 is 0.0076 mM, inhibits also isozyme APN1 Bombyx mori amastatin
0.074
-
isozyme APN4: IC50 is 0.074 mM, inhibits also isozyme APN1 Bombyx mori probestin
0.148
-
isozyme APN4: IC50 is 0.148 mM, inhibits also isozyme APN1 Bombyx mori actinonin