| IED ID | IndEnz0009000073 |
| Enzyme Type ID | chitinase000073 |
| Protein Name |
Collagenase ColG EC 3.4.24.3 Class I collagenase Gelatinase ColG Microbial collagenase |
| Gene Name | colG |
| Organism | Hathewaya histolytica (Clostridium histolyticum) |
| Taxonomic Lineage | cellular organisms Bacteria Terrabacteria group Firmicutes Clostridia Eubacteriales Clostridiaceae Hathewaya Hathewaya histolytica (Clostridium histolyticum) |
| Enzyme Sequence | MKKNILKILMDSYSKESKIQTVRRVTSVSLLAVYLTMNTSSLVLAKPIENTNDTSIKNVEKLRNAPNEENSKKVEDSKNDKVEHVKNIEEAKVEQVAPEVKSKSTLRSASIANTNSEKYDFEYLNGLSYTELTNLIKNIKWNQINGLFNYSTGSQKFFGDKNRVQAIINALQESGRTYTANDMKGIETFTEVLRAGFYLGYYNDGLSYLNDRNFQDKCIPAMIAIQKNPNFKLGTAVQDEVITSLGKLIGNASANAEVVNNCVPVLKQFRENLNQYAPDYVKGTAVNELIKGIEFDFSGAAYEKDVKTMPWYGKIDPFINELKALGLYGNITSATEWASDVGIYYLSKFGLYSTNRNDIVQSLEKAVDMYKYGKIAFVAMERITWDYDGIGSNGKKVDHDKFLDDAEKHYLPKTYTFDNGTFIIRAGDKVSEEKIKRLYWASREVKSQFHRVVGNDKALEVGNADDVLTMKIFNSPEEYKFNTNINGVSTDNGGLYIEPRGTFYTYERTPQQSIFSLEELFRHEYTHYLQARYLVDGLWGQGPFYEKNRLTWFDEGTAEFFAGSTRTSGVLPRKSILGYLAKDKVDHRYSLKKTLNSGYDDSDWMFYNYGFAVAHYLYEKDMPTFIKMNKAILNTDVKSYDEIIKKLSDDANKNTEYQNHIQELADKYQGAGIPLVSDDYLKDHGYKKASEVYSEISKAASLTNTSVTAEKSQYFNTFTLRGTYTGETSKGEFKDWDEMSKKLDGTLESLAKNSWSGYKTLTAYFTNYRVTSDNKVQYDVVFHGVLTDNADISNNKAPIAKVTGPSTGAVGRNIEFSGKDSKDEDGKIVSYDWDFGDGATSRGKNSVHAYKKAGTYNVTLKVTDDKGATATESFTIEIKNEDTTTPITKEMEPNDDIKEANGPIVEGVTVKGDLNGSDDADTFYFDVKEDGDVTIELPYSGSSNFTWLVYKEGDDQNHIASGIDKNNSKVGTFKSTKGRHYVFIYKHDSASNISYSLNIKGLGNEKLKEKENNDSSDKATVIPNFNTTMQGSLLGDDSRDYYSFEVKEEGEVNIELDKKDEFGVTWTLHPESNINDRITYGQVDGNKVSNKVKLRPGKYYLLVYKYSGSGNYELRVNK |
| Enzyme Length | 1118 |
| Uniprot Accession Number | Q9X721 |
| Absorption | |
| Active Site | ACT_SITE 524; /evidence="ECO:0000255|PROSITE-ProRule:PRU10095, ECO:0000305|PubMed:11121400" |
| Activity Regulation | ACTIVITY REGULATION: Inhibited by 1-10-phenanthroline (PubMed:18937627). Inhibited by peptidomimetic isoamyl-phosphonyl-Gly-Pro-Ala, which binds to Zn(2+) (PubMed:21947205). 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:21947205, 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:0000269|PubMed:3002446, ECO:0000305|PubMed:18937627}; |
| 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). Active on soluble type I collagen, insoluble collagen, azocoll, soluble PZ-peptide (all collagenase substrates) and gelatin (PubMed:9922257). The full-length protein has collagenase activity, while the in vivo derived C-terminally truncated shorter versions only act on gelatin (PubMed:9922257). In vitro digestion of soluble calf skin collagen fibrils requires both ColG and ColH; ColG forms missing the second collagen-binding domain are also synergistic with ColH, although their overall efficiency is decreased (PubMed:18374061, PubMed:22099748). The activator domain (residues 119-388) and catalytic subdomain (389-670) open and close around substrate using a Gly-rich hinge (387-397), allowing digestion when the protein is closed (PubMed:21947205, PubMed:23703618). Binding of collagen requires Ca(2+) and is inhibited by EGTA; the collagen-binding domain (CBD, S3a plus S3b) specifically recognizes the triple-helical conformation made by 3 collagen protein chains in the triple-helical region (PubMed:11121400). Isolated CBD (S3a plus S3b) binds collagen fibrils and sheets of many tissues (PubMed:11913772). {ECO:0000269|PubMed:11121400, ECO:0000269|PubMed:11913772, ECO:0000269|PubMed:18374061, ECO:0000269|PubMed:18937627, ECO:0000269|PubMed:21947205, ECO:0000269|PubMed:22099748, ECO:0000269|PubMed:23703618, ECO:0000269|PubMed:24125730, ECO:0000269|PubMed:28820255, ECO:0000269|PubMed:3002446, ECO:0000269|PubMed:9922257}. |
| Temperature Dependency | |
| PH Dependency | |
| Pathway | |
| nucleotide Binding | |
| Features | Active site (1); Beta strand (40); Chain (1); Domain (1); Helix (44); Metal binding (33); Mutagenesis (11); Propeptide (1); Region (9); Sequence conflict (3); Signal peptide (1); Site (4); Turn (8) |
| Keywords | 3D-structure;Calcium;Direct protein sequencing;Hydrolase;Metal-binding;Metalloprotease;Pharmaceutical;Protease;Repeat;Secreted;Signal;Virulence;Zinc;Zymogen |
| Interact With | |
| Induction | INDUCTION: RNA levels are high in late logarithmic phase. {ECO:0000269|PubMed:9922257}. |
| Subcellular Location | SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:18374061, ECO:0000269|PubMed:22099748, ECO:0000269|PubMed:9922257}. |
| Modified Residue | |
| Post Translational Modification | PTM: Upon purification gives 67 kDa, 78 kDa, 82 kDa and 116 kDa (full-length) proteins all of which have the same N-terminus; only the longest form digests insoluble collagen (PubMed:9922257). At least 2 in vivo isolated forms (C1b and C1c) are missing the second collagen-binding domain, ending on Lys-1006 and Lys-1018 respectively (PubMed:22099748). {ECO:0000269|PubMed:22099748, ECO:0000269|PubMed:9922257}. |
| Signal Peptide | SIGNAL 1..45; /evidence=ECO:0000255 |
| Structure 3D | X-ray crystallography (13) |
| Cross Reference PDB | 1NQD; 1NQJ; 2O8O; 2Y3U; 2Y50; 2Y6I; 2Y72; 4AQO; 4ARE; 4HPK; 4JRW; 4TN9; 5IKU; |
| Mapped Pubmed ID | 30035850; |
| Motif | |
| Gene Encoded By | |
| Mass | 126,242 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=0.840 mM for furylacryloyl-Leu-Gly-Pro-Ala (FALGPA) {ECO:0000269|PubMed:18937627}; Vmax=0.0852 umol/min/mg enzyme {ECO:0000269|PubMed:18937627}; Note=kcat is 0.11/sec, using a catalytic fragment (119-790) on an artificial substrate. {ECO:0000269|PubMed:18937627}; |
| Metal Binding | METAL 498; /note="Calcium 1"; /evidence="ECO:0000250|UniProtKB:Q899Y1, ECO:0000303|PubMed:23703618"; METAL 523; /note="Zinc; catalytic; via tele nitrogen"; /evidence="ECO:0000255|PROSITE-ProRule:PRU10095, ECO:0000269|PubMed:21947205, ECO:0000269|PubMed:23703618, ECO:0007744|PDB:2Y50, ECO:0007744|PDB:2Y6I, ECO:0007744|PDB:4ARE"; METAL 527; /note="Zinc; catalytic; via tele nitrogen"; /evidence="ECO:0000255|PROSITE-ProRule:PRU10095, ECO:0000269|PubMed:21947205, ECO:0000269|PubMed:23703618, ECO:0007744|PDB:2Y50, ECO:0007744|PDB:2Y6I, ECO:0007744|PDB:4ARE"; METAL 531; /note="Calcium 1; via carbonyl oxygen"; /evidence="ECO:0000250|UniProtKB:Q899Y1, ECO:0000303|PubMed:23703618"; METAL 535; /note="Calcium 1; via carbonyl oxygen"; /evidence="ECO:0000250|UniProtKB:Q899Y1, ECO:0000303|PubMed:23703618"; METAL 537; /note="Calcium 1; via carbonyl oxygen"; /evidence="ECO:0000250|UniProtKB:Q899Y1, ECO:0000303|PubMed:23703618"; METAL 555; /note="Zinc; catalytic"; /evidence="ECO:0000269|PubMed:21947205, ECO:0000269|PubMed:23703618, ECO:0007744|PDB:2Y50, ECO:0007744|PDB:2Y6I, ECO:0007744|PDB:4ARE"; METAL 795; /note="Calcium 2"; /evidence="ECO:0000269|PubMed:23703618, ECO:0007744|PDB:4AQO"; METAL 796; /note="Calcium 2; via carbonyl oxygen"; /evidence="ECO:0000269|PubMed:23703618, ECO:0007744|PDB:4AQO"; METAL 823; /note="Calcium 2"; /evidence="ECO:0000269|PubMed:23703618, ECO:0007744|PDB:4AQO"; METAL 825; /note="Calcium 2"; /evidence="ECO:0000269|PubMed:23703618, ECO:0007744|PDB:4AQO"; METAL 864; /note="Calcium 2"; /evidence="ECO:0000269|PubMed:23703618, ECO:0007744|PDB:4AQO"; METAL 890; /note="Calcium 3"; /evidence="ECO:0007744|PDB:5IKU"; METAL 892; /note="Calcium 3"; /evidence="ECO:0007744|PDB:5IKU"; METAL 892; /note="Calcium 4"; /evidence="ECO:0007744|PDB:5IKU"; METAL 894; /note="Calcium 4"; /evidence="ECO:0007744|PDB:5IKU"; METAL 913; /note="Calcium 4; via carbonyl oxygen"; /evidence="ECO:0007744|PDB:5IKU"; METAL 918; /note="Calcium 3"; /evidence="ECO:0007744|PDB:5IKU"; METAL 918; /note="Calcium 4"; /evidence="ECO:0007744|PDB:5IKU"; METAL 920; /note="Calcium 4; via carbonyl oxygen"; /evidence="ECO:0007744|PDB:5IKU"; METAL 921; /note="Calcium 3"; /evidence="ECO:0007744|PDB:5IKU"; METAL 921; /note="Calcium 4"; /evidence="ECO:0007744|PDB:5IKU"; METAL 1009; /note="Calcium 5"; /evidence="ECO:0000269|PubMed:12682007, ECO:0000269|PubMed:23144249, ECO:0007744|PDB:1NQD, ECO:0007744|PDB:2O8O, ECO:0007744|PDB:4HPK, ECO:0007744|PDB:5IKU"; METAL 1011; /note="Calcium 5"; /evidence="ECO:0000269|PubMed:12682007, ECO:0000269|PubMed:23144249, ECO:0007744|PDB:1NQD, ECO:0007744|PDB:2O8O, ECO:0007744|PDB:4HPK, ECO:0007744|PDB:5IKU"; METAL 1011; /note="Calcium 6"; /evidence="ECO:0000269|PubMed:12682007, ECO:0000269|PubMed:23144249, ECO:0007744|PDB:1NQD, ECO:0007744|PDB:2O8O, ECO:0007744|PDB:4HPK, ECO:0007744|PDB:5IKU"; METAL 1013; /note="Calcium 6"; /evidence="ECO:0000269|PubMed:12682007, ECO:0000269|PubMed:23144249, ECO:0007744|PDB:1NQD, ECO:0007744|PDB:2O8O, ECO:0007744|PDB:4HPK, ECO:0007744|PDB:5IKU"; METAL 1014; /note="Calcium 6"; /evidence="ECO:0000269|PubMed:12682007, ECO:0000269|PubMed:23144249, ECO:0007744|PDB:1NQD, ECO:0007744|PDB:2O8O, ECO:0007744|PDB:4HPK"; METAL 1032; /note="Calcium 5; via carbonyl oxygen"; /evidence="ECO:0000269|PubMed:12682007, ECO:0000269|PubMed:23144249, ECO:0007744|PDB:1NQD, ECO:0007744|PDB:2O8O, ECO:0007744|PDB:4HPK, ECO:0007744|PDB:5IKU"; METAL 1037; /note="Calcium 5"; /evidence="ECO:0000269|PubMed:12682007, ECO:0000269|PubMed:23144249, ECO:0007744|PDB:1NQD, ECO:0007744|PDB:2O8O, ECO:0007744|PDB:4HPK, ECO:0007744|PDB:5IKU"; METAL 1037; /note="Calcium 6"; /evidence="ECO:0000269|PubMed:12682007, ECO:0000269|PubMed:23144249, ECO:0007744|PDB:1NQD, ECO:0007744|PDB:2O8O, ECO:0007744|PDB:4HPK, ECO:0007744|PDB:5IKU"; METAL 1039; /note="Calcium 6; via carbonyl oxygen"; /evidence="ECO:0000269|PubMed:12682007, ECO:0000269|PubMed:23144249, ECO:0007744|PDB:1NQD, ECO:0007744|PDB:2O8O, ECO:0007744|PDB:4HPK, ECO:0007744|PDB:5IKU"; METAL 1040; /note="Calcium 5"; /evidence="ECO:0000269|PubMed:12682007, ECO:0000269|PubMed:23144249, ECO:0007744|PDB:1NQD, ECO:0007744|PDB:2O8O, ECO:0007744|PDB:4HPK, ECO:0007744|PDB:5IKU"; METAL 1040; /note="Calcium 6"; /evidence="ECO:0000269|PubMed:12682007, ECO:0000269|PubMed:23144249, ECO:0007744|PDB:1NQD, ECO:0007744|PDB:2O8O, ECO:0007744|PDB:4HPK, ECO:0007744|PDB:5IKU" |
| Rhea ID | |
| Cross Reference Brenda | 3.4.24.3; |