ID CHXA_VIBCL Reviewed; 666 AA. AC Q5EK40; DT 28-JUN-2011, integrated into UniProtKB/Swiss-Prot. DT 15-MAR-2005, sequence version 1. DT 03-MAY-2023, entry version 71. DE RecName: Full=Cholix toxin; DE EC=2.4.2.36; DE AltName: Full=Exotoxin A; DE AltName: Full=NAD(+)--diphthamide ADP-ribosyltransferase; DE Flags: Precursor; GN Name=chxA; Synonyms=toxA; OS Vibrio cholerae. OC Bacteria; Pseudomonadota; Gammaproteobacteria; Vibrionales; Vibrionaceae; OC Vibrio. OX NCBI_TaxID=666; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=TP; RX PubMed=15838025; DOI=10.1128/jb.187.9.2992-3001.2005; RA Purdy A., Rohwer F., Edwards R., Azam F., Bartlett D.H.; RT "A glimpse into the expanded genome content of Vibrio cholerae through RT identification of genes present in environmental strains."; RL J. Bacteriol. 187:2992-3001(2005). RN [2] RP X-RAY CRYSTALLOGRAPHY (1.25 ANGSTROMS) OF 459-666 IN COMPLEX WITH INHIBITOR RP AND (2.1 ANGSTROMS) OF 33-666, FUNCTION AS A TOXIN, FUNCTION AS A RP GLYCOHYDROLASE, FUNCTION AS AN ADP-RIBOSYLTRANSFERASE, BIOPHYSICOCHEMICAL RP PROPERTIES, AND MUTAGENESIS OF GLU-613. RC STRAIN=TP; RX PubMed=18276581; DOI=10.1074/jbc.m710008200; RA Jorgensen R., Purdy A.E., Fieldhouse R.J., Kimber M.S., Bartlett D.H., RA Merrill A.R.; RT "Cholix toxin, a novel ADP-ribosylating factor from Vibrio cholerae."; RL J. Biol. Chem. 283:10671-10678(2008). RN [3] RP X-RAY CRYSTALLOGRAPHY (1.19 ANGSTROMS) OF 459-665 IN COMPLEX WITH RP INHIBITOR, FUNCTION AS A TOXIN, ACTIVITY REGULATION, AND MUTAGENESIS OF RP GLU-606. RC STRAIN=TP; RX PubMed=19793133; DOI=10.1111/j.1574-6968.2009.01777.x; RA Turgeon Z., White D., Jorgensen R., Visschedyk D., Fieldhouse R.J., RA Mangroo D., Merrill A.R.; RT "Yeast as a tool for characterizing mono-ADP-ribosyltransferase toxins."; RL FEMS Microbiol. Lett. 300:97-106(2009). RN [4] RP X-RAY CRYSTALLOGRAPHY (1.27 ANGSTROMS) OF 459-665 IN COMPLEX WITH RP INHIBITORS. RC STRAIN=TP; RX PubMed=21135177; DOI=10.1128/aac.01164-10; RA Turgeon Z., Jorgensen R., Visschedyk D., Edwards P.R., Legree S., RA McGregor C., Fieldhouse R.J., Mangroo D., Schapira M., Merrill A.R.; RT "Newly discovered and characterized antivirulence compounds inhibit RT bacterial mono-ADP-ribosyltransferase toxins."; RL Antimicrob. Agents Chemother. 55:983-991(2011). CC -!- FUNCTION: An NAD-dependent ADP-ribosyltransferase (ADPRT), it catalyzes CC the transfer of the ADP-ribosyl moiety of oxidized NAD onto eukaryotic CC elongation factor 2 (eEF-2) thus arresting protein synthesis. It CC probably uses the eukaryotic prolow-density lipoprotein receptor- CC related protein 1 (LRP1) to enter mouse cells, although there seems to CC be at least one other receptor as well. Is active against mouse CC fibroblasts, Chinese hamster ovary eEF-2, brine shrimp (Artemia spp. CC nauplii) and upon expression in S.cerevisiae. CC {ECO:0000269|PubMed:18276581, ECO:0000269|PubMed:19793133}. CC -!- CATALYTIC ACTIVITY: CC Reaction=diphthamide-[translation elongation factor 2] + NAD(+) = H(+) CC + N-(ADP-D-ribosyl)diphthamide-[translation elongation factor 2] + CC nicotinamide; Xref=Rhea:RHEA:11820, Rhea:RHEA-COMP:10174, Rhea:RHEA- CC COMP:10175, ChEBI:CHEBI:15378, ChEBI:CHEBI:16692, ChEBI:CHEBI:17154, CC ChEBI:CHEBI:57540, ChEBI:CHEBI:82697; EC=2.4.2.36; CC -!- ACTIVITY REGULATION: Partially inhibited by 1,8-naphthalimide (NAP). CC {ECO:0000269|PubMed:19793133}. CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=45 uM for NAD {ECO:0000269|PubMed:18276581}; CC Note=For ADP-ribosyltransferase activity of a catalytic fragment of CC residues 459-666.; CC -!- PTM: Probably requires further proteolytic processing to generate a CC fragment with toxic activity. CC -!- MISCELLANEOUS: Exotoxins are often highly strain specific; not encoded CC in strain ATCC 39315 / El Tor Inabe N16961. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; AY876053; AAW80252.1; -; Genomic_DNA. DR PDB; 2Q5T; X-ray; 2.10 A; A=33-666. DR PDB; 2Q6M; X-ray; 1.25 A; A=459-666. DR PDB; 3ESS; X-ray; 1.19 A; A=459-665. DR PDB; 3KI0; X-ray; 1.29 A; A=459-665. DR PDB; 3KI1; X-ray; 1.43 A; A=459-665. DR PDB; 3KI2; X-ray; 1.28 A; A=459-665. DR PDB; 3KI3; X-ray; 1.27 A; A=459-665. DR PDB; 3KI4; X-ray; 1.65 A; A=459-665. DR PDB; 3KI5; X-ray; 1.55 A; A=459-665. DR PDB; 3KI6; X-ray; 1.54 A; A=459-665. DR PDB; 3KI7; X-ray; 1.32 A; A=459-665. DR PDB; 3NY6; X-ray; 1.68 A; A=459-665. DR PDB; 3Q9O; X-ray; 1.79 A; A=33-666. DR PDBsum; 2Q5T; -. DR PDBsum; 2Q6M; -. DR PDBsum; 3ESS; -. DR PDBsum; 3KI0; -. DR PDBsum; 3KI1; -. DR PDBsum; 3KI2; -. DR PDBsum; 3KI3; -. DR PDBsum; 3KI4; -. DR PDBsum; 3KI5; -. DR PDBsum; 3KI6; -. DR PDBsum; 3KI7; -. DR PDBsum; 3NY6; -. DR PDBsum; 3Q9O; -. DR AlphaFoldDB; Q5EK40; -. DR SMR; Q5EK40; -. DR DrugBank; DB08348; N~2~,N~2~-DIMETHYL-N~1~-(6-OXO-5,6-DIHYDROPHENANTHRIDIN-2-YL)GLYCINAMIDE. DR TCDB; 1.C.73.1.2; the pseudomonas exotoxin a (p-exoa) family. DR BRENDA; 2.4.2.31; 6626. DR BRENDA; 2.4.2.36; 6626. DR EvolutionaryTrace; Q5EK40; -. DR GO; GO:0047286; F:NAD+-diphthamide ADP-ribosyltransferase activity; IEA:UniProtKB-EC. DR GO; GO:0016779; F:nucleotidyltransferase activity; IEA:UniProtKB-KW. DR GO; GO:0090729; F:toxin activity; IEA:UniProtKB-KW. DR CDD; cd01436; Dipth_tox_like; 1. DR Gene3D; 2.60.120.200; -; 1. DR Gene3D; 3.90.175.10; Diphtheria Toxin, domain 1; 1. DR Gene3D; 3.90.1350.10; Exotoxin A, middle domain; 1. DR InterPro; IPR013320; ConA-like_dom_sf. DR InterPro; IPR015185; Exotox-A_bind. DR InterPro; IPR015099; Exotox-A_cataly_dom. DR InterPro; IPR015186; Exotox-A_middle_dom. DR InterPro; IPR036478; Exotox-A_middle_dom_sf. DR Pfam; PF09101; Exotox-A_bind; 1. DR Pfam; PF09009; Exotox-A_cataly; 1. DR Pfam; PF09102; Exotox-A_target; 1. DR SUPFAM; SSF56399; ADP-ribosylation; 1. DR SUPFAM; SSF49899; Concanavalin A-like lectins/glucanases; 1. DR SUPFAM; SSF56864; Exotoxin A, middle domain; 1. PE 1: Evidence at protein level; KW 3D-structure; Disulfide bond; Glycosyltransferase; NAD; KW Nucleotidyltransferase; Signal; Toxin; Transferase; Virulence. FT SIGNAL 1..32 FT /evidence="ECO:0000255" FT CHAIN 33..666 FT /note="Cholix toxin" FT /id="PRO_0000409819" FT REGION 645..666 FT /note="Disordered" FT /evidence="ECO:0000256|SAM:MobiDB-lite" FT ACT_SITE 613 FT /evidence="ECO:0000305" FT DISULFID 43..47 FT DISULFID 240..257 FT DISULFID 310..332 FT DISULFID 426..433 FT MUTAGEN 606 FT /note="E->A: Loss of ADPRT activity, loss of toxicity FT against yeast." FT /evidence="ECO:0000269|PubMed:19793133" FT MUTAGEN 613 FT /note="E->A: Loss of ADPRT activity, loss of toxicity FT against mouse cells, brine shrimp and yeast." FT /evidence="ECO:0000269|PubMed:18276581" FT HELIX 39..42 FT /evidence="ECO:0007829|PDB:3Q9O" FT STRAND 47..49 FT /evidence="ECO:0007829|PDB:3Q9O" FT STRAND 57..62 FT /evidence="ECO:0007829|PDB:3Q9O" FT STRAND 71..81 FT /evidence="ECO:0007829|PDB:3Q9O" FT STRAND 94..99 FT /evidence="ECO:0007829|PDB:3Q9O" FT TURN 100..102 FT /evidence="ECO:0007829|PDB:3Q9O" FT STRAND 103..108 FT /evidence="ECO:0007829|PDB:3Q9O" FT STRAND 113..115 FT /evidence="ECO:0007829|PDB:2Q5T" FT STRAND 120..126 FT /evidence="ECO:0007829|PDB:3Q9O" FT STRAND 131..137 FT /evidence="ECO:0007829|PDB:3Q9O" FT STRAND 139..152 FT /evidence="ECO:0007829|PDB:3Q9O" FT STRAND 157..166 FT /evidence="ECO:0007829|PDB:3Q9O" FT STRAND 170..172 FT /evidence="ECO:0007829|PDB:3Q9O" FT STRAND 177..182 FT /evidence="ECO:0007829|PDB:3Q9O" FT HELIX 185..190 FT /evidence="ECO:0007829|PDB:3Q9O" FT TURN 191..193 FT /evidence="ECO:0007829|PDB:3Q9O" FT STRAND 194..203 FT /evidence="ECO:0007829|PDB:3Q9O" FT STRAND 208..219 FT /evidence="ECO:0007829|PDB:3Q9O" FT HELIX 226..230 FT /evidence="ECO:0007829|PDB:3Q9O" FT HELIX 231..234 FT /evidence="ECO:0007829|PDB:3Q9O" FT HELIX 236..241 FT /evidence="ECO:0007829|PDB:3Q9O" FT HELIX 245..248 FT /evidence="ECO:0007829|PDB:3Q9O" FT STRAND 269..273 FT /evidence="ECO:0007829|PDB:3Q9O" FT STRAND 289..293 FT /evidence="ECO:0007829|PDB:3Q9O" FT HELIX 300..310 FT /evidence="ECO:0007829|PDB:3Q9O" FT HELIX 314..317 FT /evidence="ECO:0007829|PDB:3Q9O" FT HELIX 328..346 FT /evidence="ECO:0007829|PDB:3Q9O" FT HELIX 350..354 FT /evidence="ECO:0007829|PDB:3Q9O" FT HELIX 355..358 FT /evidence="ECO:0007829|PDB:3Q9O" FT HELIX 361..363 FT /evidence="ECO:0007829|PDB:3Q9O" FT HELIX 366..381 FT /evidence="ECO:0007829|PDB:3Q9O" FT HELIX 385..403 FT /evidence="ECO:0007829|PDB:3Q9O" FT HELIX 409..418 FT /evidence="ECO:0007829|PDB:3Q9O" FT STRAND 420..425 FT /evidence="ECO:0007829|PDB:3Q9O" FT STRAND 441..445 FT /evidence="ECO:0007829|PDB:3Q9O" FT HELIX 450..453 FT /evidence="ECO:0007829|PDB:3Q9O" FT STRAND 460..462 FT /evidence="ECO:0007829|PDB:3ESS" FT STRAND 465..468 FT /evidence="ECO:0007829|PDB:3ESS" FT HELIX 471..483 FT /evidence="ECO:0007829|PDB:3ESS" FT STRAND 486..494 FT /evidence="ECO:0007829|PDB:3ESS" FT HELIX 496..504 FT /evidence="ECO:0007829|PDB:3ESS" FT HELIX 516..520 FT /evidence="ECO:0007829|PDB:2Q6M" FT STRAND 523..529 FT /evidence="ECO:0007829|PDB:3ESS" FT HELIX 530..534 FT /evidence="ECO:0007829|PDB:3ESS" FT HELIX 545..547 FT /evidence="ECO:0007829|PDB:3ESS" FT HELIX 551..555 FT /evidence="ECO:0007829|PDB:3ESS" FT STRAND 559..566 FT /evidence="ECO:0007829|PDB:3ESS" FT HELIX 567..572 FT /evidence="ECO:0007829|PDB:3ESS" FT STRAND 573..575 FT /evidence="ECO:0007829|PDB:3ESS" FT HELIX 580..582 FT /evidence="ECO:0007829|PDB:3ESS" FT HELIX 584..591 FT /evidence="ECO:0007829|PDB:3ESS" FT STRAND 601..607 FT /evidence="ECO:0007829|PDB:3ESS" FT STRAND 612..616 FT /evidence="ECO:0007829|PDB:3ESS" FT HELIX 618..621 FT /evidence="ECO:0007829|PDB:3ESS" FT STRAND 625..632 FT /evidence="ECO:0007829|PDB:3ESS" FT HELIX 643..649 FT /evidence="ECO:0007829|PDB:3ESS" FT HELIX 650..652 FT /evidence="ECO:0007829|PDB:3ESS" FT HELIX 662..664 FT /evidence="ECO:0007829|PDB:3ESS" SQ SEQUENCE 666 AA; 74293 MW; 3A76A86E0BC7FDD7 CRC64; MYLTFYLEKV MKKMLLIAGA TVISSMAHPT FAVEDELNIF DECRSPCSLT PEPGKPIQSK LSIPSDVVLD EGVLYYSMTI NDEQNDIKDE DKGESIITIG EFATVRATRH YVNQDAPFGV IHLDITTENG TKTYSYNRKE GEFAINWLVP IGEDSPASIK ISVDELDQQR NIIEVPKLYS IDLDNQTLEQ WKTQGNVSFS VTRPEHNIAI SWPSVSYKAA QKEGSRHKRW AHWHTGLALC WLVPMDAIYN YITQQNCTLG DNWFGGSYET VAGTPKVITV KQGIEQKPVE QRIHFSKGNA MSALAAHRVC GVPLETLARS RKPRDLTDDL SCAYQAQNIV SLFVATRILF SHLDSVFTLN LDEQEPEVAE RLSDLRRINE NNPGMVTQVL TVARQIYNDY VTHHPGLTPE QTSAGAQAAD ILSLFCPDAD KSCVASNNDQ ANINIESRSG RSYLPENRAV ITPQGVTNWT YQELEATHQA LTREGYVFVG YHGTNHVAAQ TIVNRIAPVP RGNNTENEEK WGGLYVATHA EVAHGYARIK EGTGEYGLPT RAERDARGVM LRVYIPRASL ERFYRTNTPL ENAEEHITQV IGHSLPLRNE AFTGPESAGG EDETVIGWDM AIHAVAIPST IPGNAYEELA IDEEAVAKEQ SISTKPPYKE RKDELK //