| IED ID | IndEnz0002006988 |
| Enzyme Type ID | protease006988 |
| Protein Name |
Collagenase ColT EC 3.4.24.3 Microbial collagenase |
| Gene Name | colT CTC_p33 |
| Organism | Clostridium tetani (strain Massachusetts / E88) |
| Taxonomic Lineage | cellular organisms Bacteria Terrabacteria group Firmicutes Clostridia Eubacteriales Clostridiaceae Clostridium Clostridium tetani Clostridium tetani (strain Massachusetts / E88) |
| Enzyme Sequence | MKKKFIKMLCSIAIGCMISTSYSIKVSAFSNGNTKTNPNGEFKSLSLNSTNPYKTKYSFNDLNKLSNKEILDLTSKIKWSDISDLFQYNKDSYTFYSNKERVQALIDGLYEKGCNYTSTDDKGIDTLVEILRSGFYLGYYNDSLKYLNDKSFKDKCIPAMIAIENNKNFKLGENGQDTVVHALGKLIGNTSCNDEVVNKTIPILEQYYNEIDKYSKDRLKSNAVYNFMKEINYDISQYEYAHNIRDYKNTPWSGKIDSFIDTISKFASISNVTKDNGWIINNSIYYTAKLSKYHSNPSIPHSVIDNCIEIFPDYSEQYFTAIEAIKEDFNSRDSKGNVIDINKLIEEGKKHYLPKTYTFDNGKIIIKAGDKVEESKIQKLYWASKEVKSQFHRIIGNDKPLEVGNADDILTIVIYNNPEEYKLNKTLYGYSVDNGGIYIEGIGTFFTYERTPQESIYSLEELFRHEFTHYLQGRYLIPGLFNKGDFYKGNNGRITWFEEGSAEFFAGSTRTSVLPRKSMVGGLSKNPKERFNADKLLHSKYSDGWDFYKYGYAFSDYMYNNNKKLFSDLVSTMKNNDVKGYEALIEESSKDSKINKDYEYHMENLVNNYDNYTIPLVSDDYMKQYDNKSLHEIKSDIEKAMDVKNSQITKESSQYFDTYNLKATYTLSSNKGEISNWNYMNNKINEALNKLDNLSWGGYKTVTAYFSNPRLNSNNEVVYDIVFHGLLSHNKNSNEKVEVKEEPEIKDKDSFENVIYEKENNDSFDKANKIHKNQIVMATLDTEDYRDTFYFDALTSGSIDITIENIHGNSDAFNWLVYNDEDLNNYIAYPTKKEDNKLMGSFKVHKPGRYYILVYKTSLNKVNYKLNISDATNMAPVIKKIHEKENNDSFETANKITLDTLVLGNLDYKDVSDIYSFDIENTKDLNIKLTNLNNLGIAWNLYKESDLNNYIAYGAKSDNAIVGKCNLSPGKYYLYVYKYSGDKGNYSVIIN |
| Enzyme Length | 991 |
| Uniprot Accession Number | Q899Y1 |
| Absorption | |
| Active Site | ACT_SITE 466; /evidence=ECO:0000255|PROSITE-ProRule:PRU10095 |
| Activity Regulation | ACTIVITY REGULATION: Partially inhibited by 1-10-phenanthroline; inactivation is irreversible (PubMed:18937627, PubMed:23703618). Partially inhibited by EDTA; inactivation is reversible (PubMed:23703618). Inhibited by broad-spectrum zinc metalloprotease inhibitor batimastat (PubMed:28820255). N-aryl mercaptoacetamide-based inhibitors have been isolated that act on clostridial collagenases with submicromolar affinity while having negligibile activity on human collagenases (PubMed:28820255). {ECO:0000269|PubMed:18937627, ECO:0000269|PubMed:23703618, ECO:0000269|PubMed:28820255}. |
| Binding Site | |
| Calcium Binding | |
| catalytic Activity | CATALYTIC ACTIVITY: Reaction=Digestion of native collagen in the triple helical region at Xaa-|-Gly bonds. With synthetic peptides, a preference is shown for Gly at P3 and P1', Pro and Ala at P2 and P2', and hydroxyproline, Ala or Arg at P3'.; EC=3.4.24.3; Evidence={ECO:0000269|PubMed:24125730, ECO:0000305|PubMed:18937627, ECO:0000305|PubMed:23703618}; |
| DNA Binding | |
| EC Number | 3.4.24.3 |
| Enzyme Function | FUNCTION: Clostridial collagenases are among the most efficient degraders of eukaryotic collagen known; saprophytes use collagen as a carbon source while pathogens additionally digest collagen to aid in host colonization. Has both tripeptidylcarboxypeptidase on Gly-X-Y and endopeptidase activities; the endopeptidase cuts within the triple helix region of collagen while tripeptidylcarboxypeptidase successively digests the exposed ends, thus clostridial collagenases can digest large sections of collagen (By similarity). The activator domain (residues 57-330) and catalytic subdomain (340-611) open and close around substrate allowing digestion when the protein is closed (PubMed:23703618). {ECO:0000250|UniProtKB:Q9X721, ECO:0000269|PubMed:18937627, ECO:0000269|PubMed:23703618, ECO:0000269|PubMed:24125730, ECO:0000269|PubMed:28820255}. |
| Temperature Dependency | |
| PH Dependency | |
| Pathway | |
| nucleotide Binding | |
| Features | Active site (1); Beta strand (11); Chain (1); Helix (22); Metal binding (24); Propeptide (1); Region (6); Signal peptide (1); Turn (4) |
| Keywords | 3D-structure;Calcium;Hydrolase;Metal-binding;Metalloprotease;Plasmid;Protease;Reference proteome;Repeat;Secreted;Signal;Virulence;Zinc;Zymogen |
| Interact With | |
| Induction | |
| Subcellular Location | SUBCELLULAR LOCATION: Secreted {ECO:0000250|UniProtKB:Q9X721}. |
| Modified Residue | |
| Post Translational Modification | |
| Signal Peptide | SIGNAL 1..28; /evidence=ECO:0000255 |
| Structure 3D | X-ray crystallography (2) |
| Cross Reference PDB | 4AR8; 4AR9; |
| Mapped Pubmed ID | - |
| Motif | |
| Gene Encoded By | Plasmid pE88 |
| Mass | 114,376 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=0.970 mM for furylacryloyl-Leu-Gly-Pro-Ala (FALGPA) {ECO:0000269|PubMed:18937627}; Vmax=5.53 umol/min/mg enzyme {ECO:0000269|PubMed:18937627}; Note=kcat is 7.24/sec, using a catalytic fragment (residues 53-727) on an artificial substrate. {ECO:0000269|PubMed:18937627}; |
| Metal Binding | METAL 440; /note="Calcium 1"; /evidence="ECO:0000269|PubMed:23703618, ECO:0007744|PDB:4AR8, ECO:0007744|PDB:4AR9"; METAL 465; /note="Zinc; catalytic; via tele nitrogen"; /evidence="ECO:0000255|PROSITE-ProRule:PRU10095, ECO:0000269|PubMed:23703618, ECO:0007744|PDB:4AR8, ECO:0007744|PDB:4AR9"; METAL 469; /note="Zinc; catalytic; via tele nitrogen"; /evidence="ECO:0000255|PROSITE-ProRule:PRU10095, ECO:0000269|PubMed:23703618, ECO:0007744|PDB:4AR8, ECO:0007744|PDB:4AR9"; METAL 473; /note="Calcium 1; via carbonyl oxygen"; /evidence="ECO:0000269|PubMed:23703618, ECO:0007744|PDB:4AR8, ECO:0007744|PDB:4AR9"; METAL 477; /note="Calcium 1; via carbonyl oxygen"; /evidence="ECO:0000269|PubMed:23703618, ECO:0007744|PDB:4AR8, ECO:0007744|PDB:4AR9"; METAL 479; /note="Calcium 1; via carbonyl oxygen"; /evidence="ECO:0000269|PubMed:23703618, ECO:0007744|PDB:4AR8, ECO:0007744|PDB:4AR9"; METAL 499; /note="Zinc; catalytic"; /evidence="ECO:0000269|PubMed:23703618, ECO:0007744|PDB:4AR8, ECO:0007744|PDB:4AR9"; METAL 757; /note="Calcium 2"; /evidence="ECO:0000250|UniProtKB:Q9X721"; METAL 759; /note="Calcium 2"; /evidence="ECO:0000250|UniProtKB:Q9X721"; METAL 759; /note="Calcium 3"; /evidence="ECO:0000250|UniProtKB:Q9X721"; METAL 761; /note="Calcium 3"; /evidence="ECO:0000250|UniProtKB:Q9X721"; METAL 784; /note="Calcium 2"; /evidence="ECO:0000250|UniProtKB:Q9X721"; METAL 784; /note="Calcium 3"; /evidence="ECO:0000250|UniProtKB:Q9X721"; METAL 787; /note="Calcium 2"; /evidence="ECO:0000250|UniProtKB:Q9X721"; METAL 787; /note="Calcium 3"; /evidence="ECO:0000250|UniProtKB:Q9X721"; METAL 883; /note="Calcium 4"; /evidence="ECO:0000250|UniProtKB:Q9X721"; METAL 885; /note="Calcium 4"; /evidence="ECO:0000250|UniProtKB:Q9X721"; METAL 885; /note="Calcium 5"; /evidence="ECO:0000250|UniProtKB:Q9X721"; METAL 887; /note="Calcium 5"; /evidence="ECO:0000250|UniProtKB:Q9X721"; METAL 888; /note="Calcium 5"; /evidence="ECO:0000250|UniProtKB:Q9X721"; METAL 910; /note="Calcium 4"; /evidence="ECO:0000250|UniProtKB:Q9X721"; METAL 910; /note="Calcium 5"; /evidence="ECO:0000250|UniProtKB:Q9X721"; METAL 913; /note="Calcium 4"; /evidence="ECO:0000250|UniProtKB:Q9X721"; METAL 913; /note="Calcium 5"; /evidence="ECO:0000250|UniProtKB:Q9X721" |
| Rhea ID | |
| Cross Reference Brenda | 3.4.24.3; |