| IED ID | IndEnz0002002569 |
| Enzyme Type ID | protease002569 |
| Protein Name |
Multifunctional-autoprocessing repeats-in-toxin MARTX EC 3.4.22.- Cleaved into: Actin cross-linking toxin F1 EC 6.3.2.- ; Actin cross-linking toxin F4 EC 6.3.2.- ; Rho inactivation domain-containing toxin F2; ABH effector region toxin F5; Cysteine protease domain-containing toxin F3 EC 3.4.22.- |
| Gene Name | rtxA rtx VC_1451 |
| Organism | Vibrio cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961) |
| Taxonomic Lineage | cellular organisms Bacteria Proteobacteria Gammaproteobacteria Vibrionales Vibrionaceae Vibrio Vibrio cholerae Vibrio cholerae O1 Vibrio cholerae O1 biovar El Tor Vibrio cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961) |
| Enzyme Sequence | MVFYLIPKRRVWLMGKPFWRSVEYFFTGNYSADDGNNNIVAIGFGGQIHAYGGDDHVTVGSIGATVYTGSGNDTVVGGSAYLKVEDSTGHLIVKGAAGYADINKSGDGNVSFAGAAGGVSIDHLGNHGDVSYGGAAAYNGITRKGLSGNVTFAGAGGYNALWHETNQGNLSFTGAGAGNKLDRTWSNRYQGSHGDVTFDGAGAANSISSRVETGNITFRGAGADNHLVRKGKVGDITLQGAGASNRIERTHQAEDVYTQTRGNIRFEGVGGYNSLYSDVAHGDIHFSGGGAYNTIIRKGSGNDFAKEGMTNAKADEIVLTKAVMSGSWIGQDHHVTAVKSASEPNTYLFAFADSTYTKINKVQLRNDPQTGELKYYSTAWYKEVNHLSNLANQDISDNGGFTAVNINGAYTLSDLKVEHQQSVTVHAVEKSLTEYEWVTYANGAVIDAKEVSLSDAKMGGHAIYADGTKVDVKAVKSNRQPNTYIYAKVLGPYTKIVVVELANDPETGALKYQARSWYKEGDHTANIANQDISSATGYNPMGKGGYSLSDLHYSVNAVRSTSETVADIEEYTDQTLFKPANDSGESSGDVRFNGAGGGNVIKSNVTRGNVHFNGGGIANVILHSSQFGNTEFNGGGAANVIVKSGEEGDLTFRGAGLANVLVHQSEQGKMDVYAGGAVNVLVRLGDGQYLAHLLAYGNISVQKGSGDSRVVMLGGYNTHTQIGSGNGLWLAAGGFNVMTQVGKGDVAAVLAGGANVLTKMGEGELTSGMLGGANVITHISNDDQLSNTTAVALGGANILTKKGKGNTLAVMGGGANVLTHVGDGTTTGVMVGGANILTKVGNGDTTGILLGVGNVLTHVGDGQTLGVMGAAGNIFTKVGDGTSIAVMIGAGNIFTHVGEGNAWALMGGLGNVFTKVGNGDALALMVAEANVFTHIGDGMSVALMLAKGNVATKVGNGTTLAAMVGNVNIFTHIGHGSTFAAMIGQANIMTKVGNDLTAALMVGKANIMTHVGDGTSLGLFAGEVNVMTKVGNGTTLAAMFGKANIMTHVGDGLTGVLALGEANIVTKLGDDFMGVVAAAKANVVTHVGDATTAAVLAGKGNILTKVGEGTTVGLLISDVGNVMTHVGDGTTIGIAKGKANLITKVGDGLGVNVTWGQANVFTQVGDGDRYNFAKGEANLITKVGDGQEVSVVQGEANIITHVGNGDDYTGAWGKANVITKVGHGQNVVLAKGEANIVTQVGDGDSFNALWSKGNIVTKVGDGMQVTAAKGQANITTTVGNGLNVTAAYGDANINTKVGDGVSVNVAWGKYNINTKVGDGLNVAVMKGKANANIHVGDGLNINASYAQNNVAIKVGNGDFYSLAVASSNTSSNKLSALFDNIKQTVLGVGGSQAINYLVQGDEASSSGTHKGRGAIATPEITKLDGFQMDAIKEVSSDLGDSLTGSVTKVDTPDLNKMQHALNVDDSSVQAPNLIVNGDFELGEHGWQSTHGVEASYAGSVYGVEGEGHGARVTELDTYTNTSLYQDLANLAQGEVIAVSFDFAKRAGLSNNEGIEVLWNGEVVFSSSGDESAWQQKNLKLTAQAGSNRIEFKGTGHNDGLGYILDNVVATSESSQQANAIREHATQNPAAQNALSDKERAEADRQRLEQEKQKQLDAVAGSQSQLESTDQQALENNGQAQRDAVKEESEAVTAELAKLAQGLDVLDGQATHTGESGDQWRNDFAGGLLDGVQSQLDDAKQLANDKIAAAKQTLSDNNSKVKESVAKSEAGVAQGEQNRAGVEQDIADAQADAEKRKADALAKGKDAQQAESDAHHAVNNAQSRGDRDVQLAENKANQAQADAQGAKQNEGDRPDRQGVTGSGLSGNAHSVEGAGETDSHVNTDSQTNADGRFSEGLTEQEQEALEGATNAVNRLQINAGIRAKNSVSSMTSMFSETNSKSIVVPTKVSPEPERQEVTRRDVRISGVNLESLSAVQGSQPTGQLASKSVPGFKSHFASTSIGIENELSGLVVVLPKNSAQTFGYVHDSQGNPLFMLTKDMNQGGYSNPVGINDIQGVNNWQTHTIELVTYPSEISDTAAVESRKEAMLWLAKEFTDHINQSNHQSLPHLVSDDGRFTLVISNSKHLIAAGNGTSIDAQGKTIGMTPSGQQATMAISAKEFGTSSSPEVRLLESAPWYQAGLRDEFLANAKNTTLDDPATAQNVYAYLTSVYSKTADLAKEYGIYINDWDPASEGFSPNAQGLTDPKVKNAWSILPRTKPVRMLELLSAEDSRYVRQQIAEKLKGTYSESLAKNVFEYFQYGGEVAGHGINNATTGSVQQPEPAILFEFRSVPSALSDFVPKTASTVKVDVKALDHFDSASRKAIITEVNALVSGSEDFDAWYQEYRASKGQPPVKNPKSSASANHKAEWLMTQHAEQWAKITAPYTDNHETLTSTKLASNDKEELHALGETSNLENNKQQENVASIINTMLNDMLPFYALRTERNLLVQEGDEGFEVRAWPGTEDKSKTIILEDPEDAAQHKAIERFILANFDNFEQMPDELFLVDNKVISHHEGRTHVLAQKVDGAWQYNATVELMSVTELLDAANVTGKIRGESYQQVIDALTDYHASITEHADYEPESVEKLLNLRKKIEGYVLGHPDSGRVEAMNSLLNQVNTRLDEVSLLSVAEQTIQAQNSFSRLYDQLEAANLKESKHLYLDQNGDFVTKGKGNLANIDLLGSREAVLEKVKLTVSNEYGQTVADTIFAGLSAKDLAKDGKGVDIAGLNKVHQAIEQHLSPVSATLYIWKPSDHSALGHAALQIGQGRTQLEGQAAADFNQQNYVSWWPLGSKSSNISNILNVATKDQPDLKLRWSDFSQPAHQNDTLEHDVASEENDGFGLHDGDIKLKRFIEKLNAAKGIDASFKEASEGYASVLLGNPDMLETTSIPAHVFQPFVEQWNDTSYDMMDVAHRFAQELRLQAQRSDDPELLEKRIGNVIRQFAERALEEIETFKASQADQGRVFRINLEGLDVAAMQAEWHRLSNDPDARYQLLTKNCSSTVAKVLKAGGADKLIGHTWLPKFGVWTPTELFNFGQALQEAQLEIAAKKQSHQVTDVLDALSGNEKPKENVAIENDGTPPRDKESLSPLTRFLNNELYGDKEARRKIGEITQTLLDHAVEKGESQKITLQGEAGRLTGYYHQGTAPSEGETSSPSGKVVLFLHGSGSSAEEQASAIRNHYQKQGIDMLAVNLRGYGESDGGPSEKGLYQDARTMFNYLVNDKGIDPSNIIIHGYSMGGPIAADLARYAAQNGQAVSGLLLDRPMPSMTKAITAHEVANPAGIVGAIAKAVNGQFSVEKNLEGLPKETSILLLTDNEGLGNEGEKLRTKLTASGYNVTGEQTFYGHEASNRLMSQYADQIVSGLSSSASVDEDLDQQGLDTTSTKDQGISNKNDHLQVVDSKEALADGKILHNQNVNSWGPITVTPTTDGGETRFDGQIIVQMENDPVVAKAAANLAGKHAESSVVVQLDSDGNYRVVYGDPSKLDGKLRWQLVGHGRDHSETNNTRLSGYSADELAVKLAKFQQSFNQAENINNKPDHISIVGCSLVSDDKQKGFGHQFINAMDANGLRVDVSVRSSELAVDEAGRKHTKDANGDWVQKAENNKVSLSWDAQGEVVAKDERIRNGIAEGDIDLSRIGVNNVDEPARGAIGDNNDVFDAPEKRKPETEVIANSSSSNQFSYSGNIQVNVGEGEFTAVNWGTSNVGIKVGTGGFKSLAFGDNNVMVHIGDGESKHSVDIGGYQALEGAQMFLGNRNVSFNFGHSNDLILMMDKSIPTPPLVNPFDGAARISGVLQGIATSGEGEDWLAAQEQQWTLSGAKKFVKDMSGLDQSSSVDYTTLVELDSQNERDSRGLKHDAEATLNKQYNQWLSGNGNSGTSQLSRADKLRQANEKLAFNFAVGGQGADIQVTTGNWNFMFGDNIQSILDTNLGSLFGLMTQQFTATGQAKTTFTYTPQDLPRQLKNKLLGQLAGVGAETTLADIFGVDYTASGQIVSRNGQAVDGVAILKEMLEVIGEFSGDQLQAFVDPAKLLDSLKAGIDMGADGIKSFAETHGLKEKAPEEEKDNSSVSVNGANVNSAQGATVADGNTETAETQDRAFGFNSLNLPNLFATIFSQDKQKEMKSLVENLKQNLTADLLNMKEKTFDFLRNSGHLQGDGDINISLGNYNFNWGGDGKDLGAYLGDNNNFWGGRGDDVFYATGKSNIFTGGEGNDMGVLMGRENMMFGGDGNDTAVVAGRINHVFLGAGDDQSFVFGEGGEIDTGSGRDYVVTSGNFNRVDTGDDQDYSVTIGNNNQVELGAGNDFANIFGNYNRINAGAGNDVVKLMGYHAVLNGGDGDDHLIATAISKFSQFNGGEGRDLMVLGGYQNTFKGGTDVDSFVVSGDVIDNLVEDIRSEDNIVFNGIDWQKLWFERSGYDLKLSILRDPSNDSDQSKFEHIGSVTFSDYFNGNRAQVVIGMSEKDLSGEREYTMLSDSAIDALVQAMSGFEPQAGDNGFIDSLESKSQAAISMAWSDVVHKKGLMV |
| Enzyme Length | 4558 |
| Uniprot Accession Number | Q9KS12 |
| Absorption | |
| Active Site | ACT_SITE 3532; /note="For cysteine protease activity"; /evidence="ECO:0000255|PROSITE-ProRule:PRU01107, ECO:0000269|PubMed:17698571"; ACT_SITE 3581; /note="Nucleophile; for cysteine protease activity"; /evidence="ECO:0000255|PROSITE-ProRule:PRU01107, ECO:0000269|PubMed:17698571" |
| Activity Regulation | ACTIVITY REGULATION: Protease activity is inhibited by N-ethylmaleimide but not other protease inhibitors (PubMed:17464284). Protease activity is inhibited by aza-leucine epoxide (PubMed:19465933). Protease activity is activated upon binding inositol hexakisphosphate (InsP6) via an allosteric mechanism: the active site is disordered or occluded in the absence of InsP6, protecting the protease active-site sulfhydryl until the toxin enters a eukaryotic cell (PubMed:18845756, PubMed:19620709). Upon processing at the Leu-3441-Ala-3442 site, the peptidase C80 domain is converted to a form with much reduced affinity for InsP6, but is reactivated for high affinity binding of InsP6 by cooperative binding of both a new substrate and InsP6. Reactivation allows cleavage at other sites, specifically at Leu residues between the effector domains (PubMed:19620709). {ECO:0000269|PubMed:17464284, ECO:0000269|PubMed:18845756, ECO:0000269|PubMed:19465933, ECO:0000269|PubMed:19620709}. |
| Binding Site | BINDING 2255; /note="ATP; via carbonyl oxygen"; /evidence="ECO:0000250|UniProtKB:A0A0H3AIG7"; BINDING 3526; /note="Inositol hexakisphosphate"; /evidence="ECO:0000269|PubMed:18845756, ECO:0000269|PubMed:19465933, ECO:0000269|PubMed:19620709, ECO:0007744|PDB:3EEB, ECO:0007744|PDB:3FZY, ECO:0007744|PDB:3GCD"; BINDING 3577; /note="Inositol hexakisphosphate"; /evidence="ECO:0000269|PubMed:18845756, ECO:0000269|PubMed:19465933, ECO:0000269|PubMed:19620709, ECO:0007744|PDB:3EEB, ECO:0007744|PDB:3FZY, ECO:0007744|PDB:3GCD"; BINDING 3581; /note="ethyl(5S,8S,14S)-14-hydroxy-5,8,11-tris(2-methylpropyl)-3,6,9,12-tetraoxo-1-phenyl-2-oxa-4,7,10,11-tetraazapentadecan-15-oate; inhibitor; covalent"; /evidence="ECO:0000269|PubMed:19465933, ECO:0007744|PDB:3GCD"; BINDING 3636; /note="Inositol hexakisphosphate"; /evidence="ECO:0000269|PubMed:18845756, ECO:0000269|PubMed:19465933, ECO:0000269|PubMed:19620709, ECO:0007744|PDB:3EEB, ECO:0007744|PDB:3FZY, ECO:0007744|PDB:3GCD"; BINDING 3641; /note="Inositol hexakisphosphate"; /evidence="ECO:0000269|PubMed:18845756, ECO:0000269|PubMed:19465933, ECO:0000269|PubMed:19620709, ECO:0007744|PDB:3EEB, ECO:0007744|PDB:3FZY, ECO:0007744|PDB:3GCD" |
| Calcium Binding | |
| catalytic Activity | |
| DNA Binding | |
| EC Number | 3.4.22.-; 6.3.2.-; 6.3.2.-; 3.4.22.- |
| Enzyme Function | FUNCTION: [Multifunctional-autoprocessing repeats-in-toxin]: Precursor of a multifunctional toxin that causes destruction of the actin cytoskeleton by covalent cross-linking of actin and inactivation of Rho GTPases when translocated into the host cytoplasm (PubMed:26185092). Upon translocation into the host cell, undergoes autoprocessing in cis mediated by the peptidase C80 domain (also named CPD domain): the protease activity is activated upon binding inositol hexakisphosphate (InsP6) present at the host cell membrane and delivers the Cysteine protease domain-containing toxin F3 chain to the host cytosol (PubMed:17464284, PubMed:18591243, PubMed:18845756, PubMed:19620709, PubMed:19465933). The Cysteine protease domain-containing toxin F3 chain will then further cleave and release effector toxin chains that cause disassembly of the actin cytoskeleton and enhance V.cholerae colonization of the small intestine, possibly by facilitating evasion of phagocytic cells (PubMed:11553575, PubMed:12045243, PubMed:17698573, PubMed:17698571, PubMed:19812690, PubMed:19620709). {ECO:0000269|PubMed:11553575, ECO:0000269|PubMed:12045243, ECO:0000269|PubMed:17464284, ECO:0000269|PubMed:18591243, ECO:0000269|PubMed:18845756, ECO:0000269|PubMed:19465933, ECO:0000269|PubMed:19620709, ECO:0000269|PubMed:19812690, ECO:0000303|PubMed:26185092}.; FUNCTION: [Cysteine protease domain-containing toxin F3]: Following autocatalytic cleavage in cis at the Leu-3441-Ala-3442 site, this chain mediates processing in trans to release other individual toxin chains to the host cytosol (PubMed:19620709). Released effector toxin chains cause disassembly of the actin cytoskeleton and enhance V.cholerae colonization of the small intestine, possibly by facilitating evasion of phagocytic cells (PubMed:17698573, PubMed:17698571). {ECO:0000269|PubMed:17698571, ECO:0000269|PubMed:17698573, ECO:0000269|PubMed:19620709}.; FUNCTION: [Actin cross-linking toxin F1]: Actin-directed toxin that catalyzes the covalent cross-linking of host cytoplasmic monomeric actin (PubMed:11032799, PubMed:15199181, PubMed:16954226, PubMed:17951576, PubMed:19015515, PubMed:19656298, PubMed:23029200, PubMed:26228148). Mediates the cross-link between 'Lys-50' of one monomer and 'Glu-270' of another actin monomer, resulting in formation of highly toxic actin oligomers that cause cell rounding (PubMed:19015515). The toxin can be highly efficient at very low concentrations by acting on formin homology family proteins: toxic actin oligomers bind with high affinity to formins and adversely affect both nucleation and elongation abilities of formins, causing their potent inhibition in both profilin-dependent and independent manners (PubMed:26228148). Acts as an acid--amino-acid ligase that transfers the gamma-phosphoryl group of ATP to the 'Glu-270' actin residue, resulting in the formation of an activated acyl phosphate intermediate. This intermediate is further hydrolyzed and the energy of hydrolysis is utilized for the formation of the amide bond between actin subunits (PubMed:23029200). {ECO:0000269|PubMed:11032799, ECO:0000269|PubMed:15199181, ECO:0000269|PubMed:16954226, ECO:0000269|PubMed:17951576, ECO:0000269|PubMed:19015515, ECO:0000269|PubMed:19656298, ECO:0000269|PubMed:23029200, ECO:0000269|PubMed:26228148}.; FUNCTION: [Actin cross-linking toxin F4]: Actin-directed toxin that catalyzes the covalent cross-linking of host cytoplasmic monomeric actin (PubMed:11032799, PubMed:15199181, PubMed:16954226, PubMed:17951576, PubMed:19015515, PubMed:19656298, PubMed:23029200, PubMed:26228148). Mediates the cross-link between 'Lys-50' of one monomer and 'Glu-270' of another actin monomer, resulting in formation of highly toxic actin oligomers that cause cell rounding (PubMed:19015515). The toxin can be highly efficient at very low concentrations by acting on formin homology family proteins: toxic actin oligomers bind with high affinity to formins and adversely affect both nucleation and elongation abilities of formins, causing their potent inhibition in both profilin-dependent and independent manners (PubMed:26228148). Acts as an acid--amino-acid ligase that transfers the gamma-phosphoryl group of ATP to the 'Glu-270' actin residue, resulting in the formation of an activated acyl phosphate intermediate. This intermediate is further hydrolyzed and the energy of hydrolysis is utilized for the formation of the amide bond between actin subunits (PubMed:23029200). {ECO:0000269|PubMed:11032799, ECO:0000269|PubMed:15199181, ECO:0000269|PubMed:16954226, ECO:0000269|PubMed:17951576, ECO:0000269|PubMed:19015515, ECO:0000269|PubMed:19656298, ECO:0000269|PubMed:23029200, ECO:0000269|PubMed:26228148}.; FUNCTION: [Rho inactivation domain-containing toxin F2]: After delivery to the host cytosol, localizes to the host cell membrane where it modifies some cellular signaling resulting in loss of all active GTP-bound Rho and subsequent actin depolymerization. Although both this chain and the Actin cross-linking toxin F4 chain independently affect polymerized actin, the domains are not synergistic (PubMed:17474905, PubMed:19434753, PubMed:23184949). Its similarity with members of the circular permuted thiol peptidase family, suggests that it acts by mediating modification of some protein at the host cell membrane (PubMed:26185092). {ECO:0000269|PubMed:17474905, ECO:0000269|PubMed:19434753, ECO:0000269|PubMed:23184949, ECO:0000303|PubMed:26185092}.; FUNCTION: [ABH effector region toxin F5]: Indirectly activates the small GTPase CDC42. {ECO:0000269|PubMed:25427654}. |
| Temperature Dependency | |
| PH Dependency | BIOPHYSICOCHEMICAL PROPERTIES: pH dependence: Optimum pH is 7.0-9.0. {ECO:0000269|PubMed:23029200}; |
| Pathway | |
| nucleotide Binding | NP_BIND 1999..2003; /note=ATP; /evidence=ECO:0000250|UniProtKB:A0A0H3AIG7 |
| Features | Active site (2); Beta strand (13); Binding site (6); Chain (6); Compositional bias (5); Domain (2); Erroneous initiation (1); Helix (7); Metal binding (5); Mutagenesis (121); Nucleotide binding (1); Region (13); Repeat (39); Signal peptide (1); Site (4) |
| Keywords | 3D-structure;ATP-binding;Autocatalytic cleavage;Direct protein sequencing;Host cell membrane;Host cytoplasm;Host membrane;Hydrolase;Ligase;Lipid-binding;Magnesium;Membrane;Metal-binding;Multifunctional enzyme;Nucleotide-binding;Protease;Reference proteome;Repeat;Secreted;Signal;Thiol protease;Toxin;Virulence |
| Interact With | P68135 |
| Induction | |
| Subcellular Location | SUBCELLULAR LOCATION: [Multifunctional-autoprocessing repeats-in-toxin]: Secreted {ECO:0000269|PubMed:11032799, ECO:0000269|PubMed:15547287}. Host cytoplasm, host cytosol {ECO:0000303|PubMed:26185092}. Note=Secreted via the type I secretion system. {ECO:0000269|PubMed:11032799}.; SUBCELLULAR LOCATION: [Rho inactivation domain-containing toxin F2]: Host cell membrane {ECO:0000269|PubMed:20212166, ECO:0000269|PubMed:22044757}. Note=Targeted to the host cell membrane via the membrane localization region (MLD). {ECO:0000269|PubMed:20212166}.; SUBCELLULAR LOCATION: [Actin cross-linking toxin F1]: Host cytoplasm, host cytosol {ECO:0000305|PubMed:19620709}.; SUBCELLULAR LOCATION: [Actin cross-linking toxin F4]: Host cytoplasm, host cytosol {ECO:0000305|PubMed:19620709}. |
| Modified Residue | |
| Post Translational Modification | |
| Signal Peptide | SIGNAL 1..32; /evidence=ECO:0000255 |
| Structure 3D | X-ray crystallography (4) |
| Cross Reference PDB | 3EEB; 3FZY; 3GCD; 6EN3; |
| Mapped Pubmed ID | 29760474; |
| Motif | |
| Gene Encoded By | |
| Mass | 485,355 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=7.8 uM for ATP (for actin cross-linking activity) {ECO:0000269|PubMed:23029200}; KM=49.9 uM for GTP (for actin cross-linking activity) {ECO:0000269|PubMed:23029200}; |
| Metal Binding | METAL 2003; /note=Magnesium 1; catalytic; for actin cross-linking activity; /evidence=ECO:0000250|UniProtKB:A0A0H3AIG7; METAL 2003; /note=Magnesium 2; catalytic; for actin cross-linking activity; /evidence=ECO:0000250|UniProtKB:A0A0H3AIG7; METAL 2065; /note=Magnesium 2; catalytic; for actin cross-linking activity; /evidence=ECO:0000250|UniProtKB:A0A0H3AIG7; METAL 2149; /note=Magnesium 1; catalytic; for actin cross-linking activity; /evidence=ECO:0000250|UniProtKB:A0A0H3AIG7; METAL 2326; /note=Magnesium 1; catalytic; for actin cross-linking activity; /evidence=ECO:0000250|UniProtKB:A0A0H3AIG7 |
| Rhea ID | |
| Cross Reference Brenda |