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 | 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 | 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 | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
membrane | brush border | Manduca sexta | 16020 | - |
membrane | brush border | Plutella xylostella | 16020 | - |
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 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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Bombyx mori | - |
- |
- |
Drosophila melanogaster | - |
- |
- |
Epiphyas postvittana | - |
- |
- |
Helicoverpa punctigera | - |
isozymes APN1-4 | - |
Heliothis virescens | - |
- |
- |
Manduca sexta | - |
- |
- |
Plutella xylostella | - |
- |
- |
Tenebrio molitor | - |
- |
- |
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 (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 | 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 | 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 | Comment | Organism |
---|---|---|
More | the enzyme has a C-terminal anchor, and the N-terminal motif Trp-Arg-Leu-Pro | Tenebrio molitor |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
- |
Tenebrio molitor |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Tenebrio molitor | - |
- |
7.4 |
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 |