| IED ID | IndEnz0002016706 |
| Enzyme Type ID | protease016706 |
| Protein Name |
Ananain EC 3.4.22.31 |
| Gene Name | AN1 |
| Organism | Ananas comosus (Pineapple) (Ananas ananas) |
| Taxonomic Lineage | cellular organisms Eukaryota Viridiplantae Streptophyta Streptophytina Embryophyta Tracheophyta Euphyllophyta Spermatophyta Magnoliopsida Mesangiospermae Liliopsida Petrosaviidae commelinids Poales Bromeliaceae Bromelioideae Ananas Ananas comosus (Pineapple) (Ananas ananas) |
| Enzyme Sequence | MTSKVQLVFLFLFLCVMWASPSAASCDEPSDPMMKQFEEWMAEYGRVYKDNDEKMLRFQIFKNNVNHIETFNNRNGNSYTLGINQFTDMTNNEFVAQYTGLSLPLNIKREPVVSFDDVDISSVPQSIDWRDSGAVTSVKNQGRCGSCWAFASIATVESIYKIKRGNLVSLSEQQVLDCAVSYGCKGGWINKAYSFIISNKGVASAAIYPYKAAKGTCKTNGVPNSAYITRYTYVQRNNERNMMYAVSNQPIAAALDASGNFQHYKRGVFTGPCGTRLNHAIVIIGYGQDSSGKKFWIVRNSWGAGWGEGGYIRLARDVSSSFGLCGIAMDPLYPTLQSGPSVEVI |
| Enzyme Length | 345 |
| Uniprot Accession Number | P80884 |
| Absorption | |
| Active Site | ACT_SITE 147; /evidence="ECO:0000255|PROSITE-ProRule:PRU10088, ECO:0000305|PubMed:31306685"; ACT_SITE 279; /evidence="ECO:0000255|PROSITE-ProRule:PRU10089, ECO:0000305|PubMed:31306685"; ACT_SITE 300; /evidence="ECO:0000255|PROSITE-ProRule:PRU10090" |
| Activity Regulation | ACTIVITY REGULATION: Strongly inhibited by chicken egg-white cystatin (Probable). Inhibited by iodoacetamide and the active-site-directed inhibitor trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane (E-64) (PubMed:31306685). {ECO:0000269|PubMed:31306685, ECO:0000305}. |
| Binding Site | |
| Calcium Binding | |
| catalytic Activity | CATALYTIC ACTIVITY: Reaction=Hydrolysis of proteins with broad specificity for peptide bonds. Best reported small molecule substrate Bz-Phe-Val-Arg-|-NHMec, but broader specificity than fruit bromelain.; EC=3.4.22.31; Evidence={ECO:0000269|PubMed:31306685}; |
| DNA Binding | |
| EC Number | 3.4.22.31 |
| Enzyme Function | FUNCTION: Cysteine protease. Displays a high level of diversity in substrate specificity at the P1-P1' cleavage site. A hydrophilic P1 residue is preferred, with Gln or Arg strongly preferred. Favors an Ile/Leu residue at the P2 position of substrates, with an overall higher preference for Leu. The optimal tripeptide for cleavage is Pro-Leu-Gln, with cleavage occurring after the Gln residue. Another optimal tripeptide is Val-Leu-Arg, which may imply that a hydrophobic residue at the P3 position of substrates is preferred. {ECO:0000269|PubMed:31306685}. |
| Temperature Dependency | |
| PH Dependency | |
| Pathway | |
| nucleotide Binding | |
| Features | Active site (3); Beta strand (8); Chain (1); Disulfide bond (3); Helix (8); Propeptide (1); Sequence conflict (2); Signal peptide (1); Turn (1) |
| Keywords | 3D-structure;Direct protein sequencing;Disulfide bond;Hydrolase;Protease;Signal;Thiol protease;Zymogen |
| Interact With | |
| Induction | |
| Subcellular Location | |
| Modified Residue | |
| Post Translational Modification | |
| Signal Peptide | SIGNAL 1..24; /evidence=ECO:0000255 |
| Structure 3D | X-ray crystallography (6) |
| Cross Reference PDB | 6MIS; 6OKJ; 6Y6L; 6YCB; 6YCC; 6YCD; |
| Mapped Pubmed ID | 33177555; |
| Motif | |
| Gene Encoded By | |
| Mass | 38,248 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=10.5 uM for synthetic 7-amino-4-methylcoumarin (AMC) substrate Z-Phe-Val-Arg-AMC (at pH 5.0 and 37 degrees Celsius) {ECO:0000269|PubMed:31306685}; KM=18.0 uM for synthetic 7-amino-4-methylcoumarin (AMC) substrate Z-Phe-Arg-AMC (at pH 5.0 and 37 degrees Celsius) {ECO:0000269|PubMed:31306685}; KM=198 uM for synthetic 7-amino-4-methylcoumarin (AMC) substrate Z-Arg-Arg-AMC (at pH 5.0 and 37 degrees Celsius) {ECO:0000269|PubMed:31306685}; KM=5.92 uM for tripeptidyl substrate Pro-Leu-Gln (at pH 7.4 and 37 degrees Celsius) {ECO:0000269|PubMed:31306685}; KM=4.29 uM for tripeptidyl substrate Val-Leu-Arg (at pH 7.4 and 37 degrees Celsius) {ECO:0000269|PubMed:31306685}; KM=3.94 uM for tripeptidyl substrate Pro-Leu-Arg (at pH 7.4 and 37 degrees Celsius) {ECO:0000269|PubMed:31306685}; KM=4.15 uM for tripeptidyl substrate Ala-Leu-Arg (at pH 7.4 and 37 degrees Celsius) {ECO:0000269|PubMed:31306685}; KM=5.36 uM for tripeptidyl substrate Val-Leu-Lys (at pH 7.4 and 37 degrees Celsius) {ECO:0000269|PubMed:31306685}; KM=6.94 uM for tripeptidyl substrate Pro-Leu-Lys (at pH 7.4 and 37 degrees Celsius) {ECO:0000269|PubMed:31306685}; KM=5.85 uM for tripeptidyl substrate Ala-Leu-Lys (at pH 7.4 and 37 degrees Celsius) {ECO:0000269|PubMed:31306685}; KM=4.50 uM for tripeptidyl substrate Pro-Leu-Asn (at pH 7.4 and 37 degrees Celsius) {ECO:0000269|PubMed:31306685}; Note=kcat is 13.9 sec(-1) for Z-Phe-Val-Arg-AMC. kcat is 1.58 sec(-1) for Z-Phe-Arg-AMC. kcat is 0.16 sec(-1) for Z-Arg-Arg-AMC. kcat is 9.80 sec(-1) for Pro-Leu-Gln. kcat is 7.13 sec(-1) for Val-Leu-Arg. kcat is 5.62 sec(-1) for Pro-Leu-Arg. kcat is 4.96 sec(-1) for Ala-Leu-Arg. kcat is 6.29 sec(-1) for Val-Leu-Lys. kcat is 6.08 sec(-1) for Pro-Leu-Lys. kcat is 3.58 sec(-1) for Ala-Leu-Lys. kcat is 0.36 sec(-1) for Pro-Leu-Asn. {ECO:0000269|PubMed:31306685}; |
| Metal Binding | |
| Rhea ID | |
| Cross Reference Brenda |