| IED ID | IndEnz0009000286 |
| Enzyme Type ID | chitinase000286 |
| Protein Name |
Collagenase ColH EC 3.4.24.3 Class II collagenase Gelatinase ColH Microbial collagenase |
| Gene Name | colH |
| Organism | Hathewaya histolytica (Clostridium histolyticum) |
| Taxonomic Lineage | cellular organisms Bacteria Terrabacteria group Firmicutes Clostridia Eubacteriales Clostridiaceae Hathewaya Hathewaya histolytica (Clostridium histolyticum) |
| Enzyme Sequence | MKRKCLSKRLMLAITMATIFTVNSTLPIYAAVDKNNATAAVQNESKRYTVSYLKTLNYYDLVDLLVKTEIENLPDLFQYSSDAKEFYGNKTRMSFIMDEIGRRAPQYTEIDHKGIPTLVEVVRAGFYLGFHNKELNEINKRSFKERVIPSILAIQKNPNFKLGTEVQDKIVSATGLLAGNETAPPEVVNNFTPILQDCIKNIDRYALDDLKSKALFNVLAAPTYDITEYLRATKEKPENTPWYGKIDGFINELKKLALYGKINDNNSWIIDNGIYHIAPLGKLHSNNKIGIETLTEVMKVYPYLSMQHLQSADQIKRHYDSKDAEGNKIPLDKFKKEGKEKYCPKTYTFDDGKVIIKAGARVEEEKVKRLYWASKEVNSQFFRVYGIDKPLEEGNPDDILTMVIYNSPEEYKLNSVLYGYDTNNGGMYIEPEGTFFTYEREAQESTYTLEELFRHEYTHYLQGRYAVPGQWGRTKLYDNDRLTWYEEGGAELFAGSTRTSGILPRKSIVSNIHNTTRNNRYKLSDTVHSKYGASFEFYNYACMFMDYMYNKDMGILNKLNDLAKNNDVDGYDNYIRDLSSNYALNDKYQDHMQERIDNYENLTVPFVADDYLVRHAYKNPNEIYSEISEVAKLKDAKSEVKKSQYFSTFTLRGSYTGGASKGKLEDQKAMNKFIDDSLKKLDTYSWSGYKTLTAYFTNYKVDSSNRVTYDVVFHGYLPNEGDSKNSLPYGKINGTYKGTEKEKIKFSSEGSFDPDGKIVSYEWDFGDGNKSNEENPEHSYDKVGTYTVKLKVTDDKGESSVSTTTAEIKDLSENKLPVIYMHVPKSGALNQKVVFYGKGTYDPDGSIAGYQWDFGDGSDFSSEQNPSHVYTKKGEYTVTLRVMDSSGQMSEKTMKIKITDPVYPIGTEKEPNNSKETASGPIVPGIPVSGTIENTSDQDYFYFDVITPGEVKIDINKLGYGGATWVVYDENNNAVSYATDDGQNLSGKFKADKPGRYYIHLYMFNGSYMPYRINIEGSVGR |
| Enzyme Length | 1021 |
| Uniprot Accession Number | Q46085 |
| Absorption | |
| Active Site | ACT_SITE 456; /evidence="ECO:0000255|PROSITE-ProRule:PRU10095, ECO:0000305|PubMed:9452493" |
| Activity Regulation | ACTIVITY REGULATION: Inhibited by EDTA (PubMed:9452493). Inhibited by 1-10-phenanthroline (PubMed:18937627). 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:28820255, ECO:0000269|PubMed:9452493}. |
| 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:0000269|PubMed:3002446, ECO:0000305|PubMed:10217773, ECO:0000305|PubMed:18937627, ECO:0000305|PubMed:28820255}; |
| 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 (PubMed:3002446). The full-length protein has collagenase activity, while both the 116 kDa and 98 kDa forms act on gelatin (PubMed:7961400). In vitro digestion of soluble calf skin collagen fibrils requires both ColG and ColH; ColG forms missing the second collagen-binding domain is also synergistic with ColH, although their overall efficiency is decreased (PubMed:18374061, PubMed:22099748). Digestion of collagen requires Ca(2+) and is inhibited by EDTA (PubMed:9452493). The activator domain (residues 119-388) and catalytic subdomain (330-601) open and close around substrate allowing digestion when the protein is closed (PubMed:23703618). {ECO:0000269|PubMed:18374061, ECO:0000269|PubMed:18937627, ECO:0000269|PubMed:22099748, ECO:0000269|PubMed:23703618, ECO:0000269|PubMed:24125730, ECO:0000269|PubMed:28820255, ECO:0000269|PubMed:3002446, ECO:0000269|PubMed:7961400, ECO:0000269|PubMed:9452493}. |
| Temperature Dependency | |
| PH Dependency | |
| Pathway | |
| nucleotide Binding | |
| Features | Active site (1); Beta strand (33); Chain (1); Domain (2); Helix (23); Metal binding (29); Mutagenesis (12); Propeptide (1); Region (9); Signal peptide (1); Site (1); Turn (2) |
| Keywords | 3D-structure;Calcium;Direct protein sequencing;Hydrolase;Metal-binding;Metalloprotease;Pharmaceutical;Protease;Repeat;Secreted;Signal;Virulence;Zinc;Zymogen |
| Interact With | |
| Induction | |
| Subcellular Location | SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:18374061, ECO:0000269|PubMed:22099748, ECO:0000269|PubMed:7961400}. |
| Modified Residue | |
| Post Translational Modification | PTM: Upon purification gives rise to 98 kDa, 105 kDa and 116 kDa (full-length) proteins, all of which have the same N-terminus (PubMed:7961400, PubMed:9922257). {ECO:0000269|PubMed:7961400, ECO:0000269|PubMed:9922257}. |
| Signal Peptide | SIGNAL 1..30; /evidence=ECO:0000255 |
| Structure 3D | X-ray crystallography (8) |
| Cross Reference PDB | 3JQW; 3JQX; 4AR1; 4ARF; 4JGU; 4U6T; 4U7K; 5O7E; |
| Mapped Pubmed ID | - |
| Motif | |
| Gene Encoded By | |
| Mass | 116,377 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=0.88 mM for Pz peptide (4-phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-D-Arg) {ECO:0000269|PubMed:10217773}; KM=0.269 mM for furylacryloyl-Leu-Gly-Pro-Ala (FALGPA) with a catalytic fragment (residues 41-717) {ECO:0000269|PubMed:18937627}; KM=62 uM for (7-Methoxycoumarin-4-yl)acetyl-Ala-Gly-Pro-Pro-Gly-Pro-N-3-(2,4-dinitrophenyl)-L-2,3-diaminopropionyl)-Gly-Arg-NH2 with peptidase fragment (residues 331-721) {ECO:0000269|PubMed:28820255}; Vmax=12.1 umol/min/mg enzyme on FALGPA with a catalytic fragment (residues 41-717) {ECO:0000269|PubMed:18937627}; Note=kcat is 0.11 per second on Pz peptide with whole enzyme (PubMed:10217773). kcat is 15.9 per second on FALGPA with a catalytic fragment (residues 41-717) (PubMed:18937627). {ECO:0000269|PubMed:10217773, ECO:0000269|PubMed:18937627}; |
| Metal Binding | METAL 421; /note="Zinc; catalytic"; /evidence="ECO:0000269|PubMed:23703618, ECO:0007744|PDB:4AR1"; METAL 430; /note="Calcium 1"; /evidence="ECO:0000269|PubMed:23703618, ECO:0000269|PubMed:28820255, ECO:0007744|PDB:4AR1, ECO:0007744|PDB:4ARF, ECO:0007744|PDB:5O7E"; METAL 455; /note="Zinc; catalytic; via tele nitrogen"; /evidence="ECO:0000255|PROSITE-ProRule:PRU10095, ECO:0000269|PubMed:23703618, ECO:0000269|PubMed:28820255, ECO:0000305|PubMed:10217773, ECO:0007744|PDB:4AR1, ECO:0007744|PDB:4ARF, ECO:0007744|PDB:5O7E"; METAL 459; /note="Zinc; catalytic; via tele nitrogen"; /evidence="ECO:0000255|PROSITE-ProRule:PRU10095, ECO:0000269|PubMed:23703618, ECO:0000269|PubMed:28820255, ECO:0000305|PubMed:10217773, ECO:0007744|PDB:4AR1, ECO:0007744|PDB:4ARF, ECO:0007744|PDB:5O7E"; METAL 463; /note="Calcium 1; via carbonyl oxygen"; /evidence="ECO:0000269|PubMed:23703618, ECO:0000269|PubMed:28820255, ECO:0007744|PDB:4AR1, ECO:0007744|PDB:4ARF, ECO:0007744|PDB:5O7E"; METAL 467; /note="Calcium 1; via carbonyl oxygen"; /evidence="ECO:0000269|PubMed:23703618, ECO:0000269|PubMed:28820255, ECO:0007744|PDB:4AR1, ECO:0007744|PDB:4ARF, ECO:0007744|PDB:5O7E"; METAL 469; /note="Calcium 1; via carbonyl oxygen"; /evidence="ECO:0000269|PubMed:23703618, ECO:0000269|PubMed:28820255, ECO:0007744|PDB:4AR1, ECO:0007744|PDB:4ARF, ECO:0007744|PDB:5O7E"; METAL 487; /note="Zinc; catalytic"; /evidence="ECO:0000269|PubMed:23703618, ECO:0000269|PubMed:28820255, ECO:0000305|PubMed:10217773, ECO:0007744|PDB:4AR1, ECO:0007744|PDB:4ARF, ECO:0007744|PDB:5O7E"; METAL 725; /note="Calcium 2"; /evidence="ECO:0000269|PubMed:25760606, ECO:0007744|PDB:4U7K"; METAL 726; /note="Calcium 2; via carbonyl oxygen"; /evidence="ECO:0000269|PubMed:25760606, ECO:0007744|PDB:4U7K"; METAL 753; /note="Calcium 2"; /evidence="ECO:0000269|PubMed:25760606, ECO:0007744|PDB:4U7K"; METAL 755; /note="Calcium 2"; /evidence="ECO:0000269|PubMed:25760606, ECO:0007744|PDB:4U7K"; METAL 794; /note="Calcium 2"; /evidence="ECO:0000269|PubMed:25760606, ECO:0007744|PDB:4U7K"; METAL 814; /note="Calcium 3"; /evidence="ECO:0000269|PubMed:25760606, ECO:0007744|PDB:4JGU"; METAL 815; /note="Calcium 3; via carbonyl oxygen"; /evidence="ECO:0000269|PubMed:25760606, ECO:0007744|PDB:4JGU"; METAL 842; /note="Calcium 3"; /evidence="ECO:0000269|PubMed:25760606, ECO:0007744|PDB:4JGU"; METAL 844; /note="Calcium 3"; /evidence="ECO:0000269|PubMed:25760606, ECO:0007744|PDB:4JGU"; METAL 884; /note="Calcium 3"; /evidence="ECO:0000269|PubMed:25760606, ECO:0007744|PDB:4JGU"; METAL 908; /note="Calcium 4"; /evidence="ECO:0000269|PubMed:23144249, ECO:0007744|PDB:3JQW"; METAL 910; /note="Calcium 4"; /evidence="ECO:0000269|PubMed:23144249, ECO:0007744|PDB:3JQW"; METAL 910; /note="Calcium 5"; /evidence="ECO:0000269|PubMed:23144249, ECO:0007744|PDB:3JQW, ECO:0007744|PDB:3JQX"; METAL 912; /note="Calcium 5"; /evidence="ECO:0000269|PubMed:23144249, ECO:0007744|PDB:3JQW, ECO:0007744|PDB:3JQX"; METAL 913; /note="Calcium 5"; /evidence="ECO:0000269|PubMed:23144249, ECO:0007744|PDB:3JQW, ECO:0007744|PDB:3JQX"; METAL 931; /note="Calcium 4; via carbonyl oxygen"; /evidence="ECO:0000269|PubMed:23144249, ECO:0007744|PDB:3JQW"; METAL 937; /note="Calcium 4"; /evidence="ECO:0000269|PubMed:23144249, ECO:0007744|PDB:3JQW"; METAL 937; /note="Calcium 5"; /evidence="ECO:0000269|PubMed:23144249, ECO:0007744|PDB:3JQW, ECO:0007744|PDB:3JQX"; METAL 938; /note="Calcium 5; via carbonyl oxygen"; /evidence="ECO:0000269|PubMed:23144249, ECO:0007744|PDB:3JQW, ECO:0007744|PDB:3JQX"; METAL 939; /note="Calcium 4"; /evidence="ECO:0000269|PubMed:23144249, ECO:0007744|PDB:3JQW"; METAL 939; /note="Calcium 5"; /evidence="ECO:0000269|PubMed:23144249, ECO:0007744|PDB:3JQW, ECO:0007744|PDB:3JQX" |
| Rhea ID | |
| Cross Reference Brenda | 3.4.24.3; |