| IED ID | IndEnz0002013059 |
| Enzyme Type ID | protease013059 |
| Protein Name |
Cytotoxin-L EC 3.4.22.- Lethal toxin LT Cleaved into: Glucosyltransferase TcsL EC 2.4.1.- |
| Gene Name | tcsL H477_0263 |
| Organism | Paeniclostridium sordellii (strain ATCC 9714 / DSM 2141 / JCM 3814 / LMG 15708 / NCIMB 10717 / 211) (Clostridium sordellii) |
| Taxonomic Lineage | cellular organisms Bacteria Terrabacteria group Firmicutes Clostridia Eubacteriales Peptostreptococcaceae Paeniclostridium Paeniclostridium sordellii (Clostridium sordellii) Paeniclostridium sordellii (strain ATCC 9714 / DSM 2141 / JCM 3814 / LMG 15708 / NCIMB 10717 / 211) (Clostridium sordellii) |
| Enzyme Sequence | MNLVNKAQLQKMAYVKFRIQEDEYVAILNALEEYHNMSESSVVEKYLKLKDINNLTDNYLNTYKKSGRNKALKKFKEYLTMEVLELKNNSLTPVEKNLHFIWIGGQINDTAINYINQWKDVNSDYTVKVFYDSNAFLINTLKKTIVESATNNTLESFRENLNDPEFDYNKFYRKRMEIIYDKQKHFIDYYKSQIEENPEFIIDNIIKTYLSNEYSKDLEALNKYIEESLNKITANNGNDIRNLEKFADEDLVRLYNQELVERWNLAAASDILRISMLKEDGGVYLDVDMLPGIQPDLFKSINKPDSITNTSWEMIKLEAIMKYKEYIPGYTSKNFDMLDEEVQRSFESALSSKSDKSEIFLPLDDIKVSPLEVKIAFANNSVINQALISLKDSYCSDLVINQIKNRYKILNDNLNPSINEGTDFNTTMKIFSDKLASISNEDNMMFMIKITNYLKVGFAPDVRSTINLSGPGVYTGAYQDLLMFKDNSTNIHLLEPELRNFEFPKTKISQLTEQEITSLWSFNQARAKSQFEEYKKGYFEGALGEDDNLDFAQNTVLDKDYVSKKILSSMKTRNKEYIHYIVQLQGDKISYEASCNLFSKDPYSSILYQKNIEGSETAYYYSVADAEIKEIDKYRIPYQISNKRKIKLTFIGHGKSEFNTDTFANLDVDSLSSEIETILNLAKADISPKYIEINLLGCNMFSYSISAEETYPGKLLLKIKDRVSELMPSISQDSITVSANQYEVRINEEGKREILDHSGKWINKEESIIKDISSKEYISFNPKENKIIVKSKYLHELSTLLQEIRNNANSSDIDLEKKVMLTECEINVASNIDRQIVEGRIEEAKNLTSDSINYIKNEFKLIESISDSLYDLKHQNGLDDSHFISFEDISKTENGFRIRFINKETGNSIFIETEKEIFSEYATHISKEISNIKDTIFDNVNGKLVKKVNLDAAHEVNTLNSAFFIQSLIEYNTTKESLSNLSVAMKVQVYAQLFSTGLNTITDASKVVELVSTALDETIDLLPTLSEGLPIIATIIDGVSLGAAIKELSETNDPLLRQEIEAKIGIMAVNLTAASTAIVTSALGIASGFSILLVPLAGISAGIPSLVNNELILQDKATKVIDYFKHISLAETEGAFTLLDDKIIMPQDDLVLSEIDFNNNSITLGKCEIWRAEGGSGHTLTDDIDHFFSSPSITYRKPWLSIYDVLNIKKEKIDFSKDLMVLPNAPNRVFGYEMGWTPGFRSLDNDGTKLLDRIRDHYEGQFYWRYFAFIADALITKLKPRYEDTNVRINLDGNTRSFIVPVITTEQIRKNLSYSFYGSGGSYSLSLSPYNMNIDLNLVENDTWVIDVDNVVKNITIESDEIQKGELIENILSKLNIEDNKIILNNHTINFYGDINESNRFISLTFSILEDINIIIEIDLVSKSYKILLSGNCMKLIENSSDIQQKIDHIGFNGEHQKYIPYSYIDNETKYNGFIDYSKKEGLFTAEFSNESIIRNIYMPDSNNLFIYSSKDLKDIRIINKGDVKLLIGNYFKDDMKVSLSFTIEDTNTIKLNGVYLDENGVAQILKFMNNAKSALNTSNSLMNFLESINIKNIFYNNLDPNIEFILDTNFIISGSNSIGQFELICDKDKNIQPYFIKFKIKETSYTLYVGNRQNLIVEPSYHLDDSGNISSTVINFSQKYLYGIDRYVNKVIIAPNLYTDEINITPVYKPNYICPEVIILDANYINEKINVNINDLSIRYVWDNDGSDLILIANSEEDNQPQVKIRFVNVFKSDTAADKLSFNFSDKQDVSVSKIISTFSLAAYSDGFFDYEFGLVSLDNDYFYINSFGNMVSGLIYINDSLYYFKPPKNNLITGFTTIDGNKYYFDPTKSGAASIGEITIDGKDYYFNKQGILQVGVINTSDGLKYFAPAGTLDENLEGESVNFIGKLNIDGKIYYFEDNYRAAVEWKLLDDETYYFNPKTGEALKGLHQIGDNKYYFDDNGIMQTGFITINDKVFYFNNDGVMQVGYIEVNGKYFYFGKNGERQLGVFNTPDGFKFFGPKDDDLGTEEGELTLYNGILNFNGKIYFFDISNTAVVGWGTLDDGSTYYFDDNTAEACIGLTVINDCKYYFDDNGIRQLGFITINDNIFYFSESGKIELGYQNINGNYFYIDESGLVLIGVFDTPDGYKYFAPLNTVNDNIYGQAVKYSGLVRVNEDVYYFGETYKIETGWIENETDKYYFDPETKKAYKGINVVDDIKYYFDENGIMRTGLISFENNNYYFNEDGKMQFGYLNIKDKMFYFGKDGKMQIGVFNTPDGFKYFAHQNTLDENFEGESINYTGWLDLDGKRYYFTDEYIAATGSLTIDGYNYYFDPDTAELVVSE |
| Enzyme Length | 2364 |
| Uniprot Accession Number | T0D3N5 |
| Absorption | |
| Active Site | ACT_SITE 653; /note=For protease activity; /evidence=ECO:0000255|PROSITE-ProRule:PRU01107; ACT_SITE 698; /note=Nucleophile; for protease activity; /evidence=ECO:0000255|PROSITE-ProRule:PRU01107 |
| Activity Regulation | ACTIVITY REGULATION: [Cytotoxin-L]: Protease activity is activated upon binding to 1D-myo-inositol hexakisphosphate (InsP6), which induces conformational reorganization. {ECO:0000250|UniProtKB:Q46342}. |
| Binding Site | BINDING 139; /note=UDP-alpha-D-glucose; /evidence=ECO:0000250|UniProtKB:Q46342; BINDING 577; /note=1D-myo-inositol hexakisphosphate; /evidence=ECO:0000250|UniProtKB:P16154; BINDING 600; /note=1D-myo-inositol hexakisphosphate; /evidence=ECO:0000250|UniProtKB:P16154; BINDING 647; /note=1D-myo-inositol hexakisphosphate; /evidence=ECO:0000250|UniProtKB:P16154; BINDING 764; /note=1D-myo-inositol hexakisphosphate; /evidence=ECO:0000250|UniProtKB:P16154; BINDING 775; /note=1D-myo-inositol hexakisphosphate; /evidence=ECO:0000250|UniProtKB:P16154; BINDING 792; /note=1D-myo-inositol hexakisphosphate; /evidence=ECO:0000250|UniProtKB:P16154 |
| Calcium Binding | |
| catalytic Activity | CATALYTIC ACTIVITY: [Glucosyltransferase TcsL]: Reaction=L-threonyl-[protein] + UDP-alpha-D-glucose = 3-O-(alpha-D-glucosyl)-L-threonyl-[protein] + H(+) + UDP; Xref=Rhea:RHEA:64684, Rhea:RHEA-COMP:11060, Rhea:RHEA-COMP:16656, ChEBI:CHEBI:15378, ChEBI:CHEBI:30013, ChEBI:CHEBI:58223, ChEBI:CHEBI:58885, ChEBI:CHEBI:156085; Evidence={ECO:0000250|UniProtKB:Q46342};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:64685; Evidence={ECO:0000250|UniProtKB:Q46342}; |
| DNA Binding | |
| EC Number | 3.4.22.-; 2.4.1.- |
| Enzyme Function | FUNCTION: [Cytotoxin-L]: Precursor of a cytotoxin that targets the vascular endothelium, inducing an anti-inflammatory effect and resulting in lethal toxic shock syndrome (PubMed:19527792, PubMed:21199912). TcsL constitutes the main toxin that mediates the pathology of P.sordellii infection, an anaerobic Gram-positive bacterium found in soil and in the gastrointestinal and vaginal tracts of animals and humans; although the majority of carriers are asymptomatic, pathogenic P.sordellii infections arise rapidly and are highly lethal (By similarity). This form constitutes the precursor of the toxin: it enters into host cells and mediates autoprocessing to release the active toxin (Glucosyltransferase TcsL) into the host cytosol (By similarity). Targets vascular endothelium by binding to the semaphorin proteins SEMA6A and SEMA6B, and enters host cells via clathrin-mediated endocytosis (By similarity). Once entered into host cells, acidification in the endosome promotes the membrane insertion of the translocation region and formation of a pore, leading to translocation of the GT44 and peptidase C80 domains across the endosomal membrane (By similarity) (PubMed:11500421). This activates the peptidase C80 domain and autocatalytic processing, releasing the N-terminal part (Glucosyltransferase TcsL), which constitutes the active part of the toxin, in the cytosol (By similarity). {ECO:0000250|UniProtKB:P18177, ECO:0000250|UniProtKB:Q46342, ECO:0000269|PubMed:11500421, ECO:0000269|PubMed:19527792, ECO:0000269|PubMed:21199912}.; FUNCTION: [Glucosyltransferase TcsL]: Active form of the toxin, which is released into the host cytosol following autoprocessing and inactivates small GTPases. Acts by mediating monoglucosylation of small GTPases of the Ras (H-Ras/HRAS, K-Ras/KRAS and N-Ras/NRAS) family in host cells at the conserved threonine residue located in the switch I region ('Thr-37/35'), using UDP-alpha-D-glucose as the sugar donor. Also able to catalyze monoglucosylation of some members of the Rho family (Rac1 and Rap2A), but with less efficiency than with Ras proteins. Monoglucosylation of host small GTPases completely prevents the recognition of the downstream effector, blocking the GTPases in their inactive form and leading to apoptosis. Induces an anti-inflammatory effect, mainly by inactivating Ras proteins which results in blockage of the cell cycle and killing of immune cells. The absence or moderate local inflammatory response allows C.sordellii spreading in deep tissues, production of toxin which is released in the general circulation and causes a toxic shock syndrome. {ECO:0000250|UniProtKB:Q46342}. |
| Temperature Dependency | |
| PH Dependency | |
| Pathway | |
| nucleotide Binding | |
| Features | Active site (2); Binding site (7); Chain (2); Domain (2); Metal binding (7); Region (14); Repeat (20); Site (1) |
| Keywords | Autocatalytic cleavage;Glycosyltransferase;Host cell membrane;Host cytoplasm;Host endosome;Host membrane;Hydrolase;Lipid-binding;Magnesium;Manganese;Membrane;Metal-binding;Protease;Reference proteome;Repeat;Secreted;Thiol protease;Toxin;Transferase;Virulence;Zinc |
| Interact With | |
| Induction | |
| Subcellular Location | SUBCELLULAR LOCATION: [Cytotoxin-L]: Secreted {ECO:0000250|UniProtKB:P18177}. Host endosome membrane {ECO:0000250|UniProtKB:P18177}. Note=Secreted from P.sordellii cell into the extracellular environment via help of holin-like protein TcdE/UtxA. Binds to the cell surface receptors via the receptor-binding region and enters the cells via clathrin-mediated endocytosis. Acidification in the endosome triggers conformational changes that promote the membrane insertion of the translocation region, allowing formation of a pore, leading to translocation of the GT44 and peptidase C80 domains across the endosomal membrane (By similarity). 1D-myo-inositol hexakisphosphate-binding (InsP6) activates the peptidase C80 domain and autoprocessing, generating the Glucosyltransferase TcsL form, which is released in the host cytosol (By similarity). {ECO:0000250|UniProtKB:P18177, ECO:0000250|UniProtKB:Q46342}.; SUBCELLULAR LOCATION: [Glucosyltransferase TcsL]: Host cytoplasm, host cytosol {ECO:0000250|UniProtKB:Q46342}. Host cell membrane {ECO:0000250|UniProtKB:Q46342}; Peripheral membrane protein {ECO:0000250|UniProtKB:Q46342}; Cytoplasmic side {ECO:0000250|UniProtKB:Q46342}. Note=Binding to phospholipids, such as phosphatidylserine and phosphatidic acid promotes localization to the inner face of the cell membrane close to its membrane anchored substrates (small GTPases). {ECO:0000250|UniProtKB:Q46342}. |
| Modified Residue | |
| Post Translational Modification | PTM: [Cytotoxin-L]: Undergoes autocatalytic cleavage to release the N-terminal part (Glucosyltransferase TcsL), which constitutes the active part of the toxin, in the host cytosol. 1D-myo-inositol hexakisphosphate-binding (InsP6) activates the peptidase C80 domain and promotes autoprocessing. {ECO:0000250|UniProtKB:Q46342}. |
| Signal Peptide | |
| Structure 3D | |
| Cross Reference PDB | - |
| Mapped Pubmed ID | - |
| Motif | |
| Gene Encoded By | |
| Mass | 270,466 |
| Kinetics | |
| Metal Binding | METAL 288; /note=Magnesium or manganese; /evidence=ECO:0000250|UniProtKB:Q46342; METAL 515; /note=Magnesium or manganese; /evidence=ECO:0000250|UniProtKB:Q46342; METAL 518; /note=Magnesium or manganese; /evidence=ECO:0000250|UniProtKB:Q46342; METAL 545; /note=Zinc; via carbonyl oxygen; /evidence=ECO:0000250|UniProtKB:P16154; METAL 546; /note=Zinc; /evidence=ECO:0000250|UniProtKB:P18177; METAL 653; /note=Zinc; /evidence=ECO:0000250|UniProtKB:P18177; METAL 757; /note=Zinc; /evidence=ECO:0000250|UniProtKB:P18177 |
| Rhea ID | RHEA:64684; RHEA:64685 |
| Cross Reference Brenda |