Detail Information for IndEnz0002016706
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