| IED ID | IndEnz0002011930 |
| Enzyme Type ID | protease011930 |
| Protein Name |
Thrombin-like enzyme collinein-1 SVTLE collinein-1 EC 3.4.21.- Fibrinogen-clotting enzyme Snake venom serine protease SVSP |
| Gene Name | |
| Organism | Crotalus durissus collilineatus (Brazilian rattlesnake) |
| Taxonomic Lineage | cellular organisms Eukaryota Opisthokonta Metazoa Eumetazoa Bilateria Deuterostomia Chordata Craniata Vertebrata Gnathostomata (jawed vertebrates) Teleostomi Euteleostomi Sarcopterygii Dipnotetrapodomorpha Tetrapoda Amniota Sauropsida Sauria (diapsids) Lepidosauria (lepidosaurs) Squamata (squamates) Bifurcata (split-tongued squamates) Unidentata Episquamata Toxicofera Serpentes (snakes) Colubroidea Viperidae Crotalinae (pit vipers) Crotalus Crotalus durissus (tropical rattlesnake) Crotalus durissus collilineatus (Brazilian rattlesnake) |
| Enzyme Sequence | VIGGDECNINEHNFLVALYEYWSQSFLCGGTLINGEWVLTAAHCDRKHILIYVGVHDRSVQFDKEQRRFPKEKYFFNCRNNFTKWDKDIMLIRLNKPVSYSEHIAPLSLPSSPPIVGSVCRVMGWGTIKSPQETLPDVPHCANINLLDYEVCRTAHPQFRLPATIRILCAGVLEGGIDTCHRDSGGPLICNGEFQGIVSWGDGSCAQPDKPALYSKVFDHLDWIQNIIAGSETVNCPS |
| Enzyme Length | 238 |
| Uniprot Accession Number | A0A0S4FKT4 |
| Absorption | |
| Active Site | ACT_SITE 43; /note=Charge relay system; /evidence=ECO:0000255|PROSITE-ProRule:PRU00274; ACT_SITE 88; /note=Charge relay system; /evidence=ECO:0000255|PROSITE-ProRule:PRU00274; ACT_SITE 184; /note=Charge relay system; /evidence=ECO:0000255|PROSITE-ProRule:PRU00274 |
| Activity Regulation | ACTIVITY REGULATION: Inhibited by Cu(2+) and, to a lesser extent, by Zn(2+) and Ba(2+). Not inhibited by Ca(2+) and Mg(2+). {ECO:0000269|PubMed:26227411}. |
| Binding Site | |
| Calcium Binding | |
| catalytic Activity | |
| DNA Binding | |
| EC Number | 3.4.21.- |
| Enzyme Function | FUNCTION: Thrombin-like snake venom serine protease (PubMed:26227411, PubMed:31131001, PubMed:34506860). Releases fibrinopeptide A and B in the conversion of fibrinogen to fibrin, with preferential activity on the alpha chain of fibrinogen (PubMed:26227411, PubMed:34506860). Also hydrolyzes N-p-toluensulfonyl arginine ester (TAME) and chromogenic artificial substrates of the blood coagulation cascade: S-2222 for factor Xa, S-2302 for kallikrein and S-2238 for thrombin (PubMed:26227411). When tested in vitro, the recombinant protein does not degrade blood clots, suggesting that this toxin lacks fibrinolytic activity (PubMed:31131001). In addition, it moderately inhibits human Kv10.1/KCNH1/EAG1 currents, with a mechanism independent of its enzymatic activity. It selectively blocks Kv10.1/KCNH1/EAG1 in a time and dose-dependent manner (IC(50)=4.2 uM for native protein and IC(50)=2.5 uM for recombinant protein). It may have a preference in interacting with Kv10.1/KCNH1/EAG1 in its closed state, since the inhibitory effect of the toxin is decreased at more depolarized potentials. Corroboratively, it may have possible antitumor applications, since it reduces the viability of human breast cancer cell line MCF7, which strongly expresses Kv10.1/KCNH1/EAG1, but does not affect the liver carcinoma and the non-tumorigenic epithelial breast cell lines, which weakly express Kv10.1/KCNH1/EAG1 (PubMed:32161292). When tested on peripheral blood mononuclear cells (PBMC), the native protein shows mild cytotoxicity, whereas the recombinant protein does not show any cytotoxicity (PubMed:34506860). Native form is not immununogenic, since it does not induce statistically significant antibody production in mice, whereas recombinant form shows an antibody titer slightly higher than the native form (PubMed:34506860). In vivo, subplantar injection in mice paw induces a discreet paw edema (PubMed:31131001). In addition, intraperitoneal injection of the recombinant protein into mice led to fibrinogen depletion, resulting in the blood incoagulability (PubMed:31131001). {ECO:0000269|PubMed:26227411, ECO:0000269|PubMed:31131001, ECO:0000269|PubMed:32161292, ECO:0000269|PubMed:34506860}. |
| Temperature Dependency | BIOPHYSICOCHEMICAL PROPERTIES: Temperature dependence: Thermostable. Activity is stable after incubation at 100 degrees Celsius for 30 min. {ECO:0000269|PubMed:26227411}; |
| PH Dependency | BIOPHYSICOCHEMICAL PROPERTIES: pH dependence: Activity is stable from pH 3.5 and 10.5. {ECO:0000269|PubMed:26227411}; |
| Pathway | |
| nucleotide Binding | |
| Features | Active site (3); Chain (1); Disulfide bond (6); Domain (1); Mutagenesis (1); Site (1) |
| Keywords | Blood coagulation cascade activating toxin;Direct protein sequencing;Disulfide bond;Hemostasis impairing toxin;Hydrolase;Ion channel impairing toxin;Potassium channel impairing toxin;Protease;Secreted;Serine protease;Toxin;Voltage-gated potassium channel impairing toxin |
| Interact With | |
| Induction | |
| Subcellular Location | SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:26227411}. |
| Modified Residue | |
| Post Translational Modification | |
| Signal Peptide | |
| Structure 3D | |
| Cross Reference PDB | - |
| Mapped Pubmed ID | - |
| Motif | |
| Gene Encoded By | |
| Mass | 26,664 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=1.43 mM for TAME {ECO:0000269|PubMed:26227411}; KM=0.92 mM for S-2302 {ECO:0000269|PubMed:34506860}; Vmax=0.06 mmol/min/ug enzyme towards TAME {ECO:0000269|PubMed:26227411}; |
| Metal Binding | |
| Rhea ID | |
| Cross Reference Brenda |