CPNT_MYCTU
ID CPNT_MYCTU Reviewed; 846 AA.
AC O05442; F2GDR4; I6Y4T3; Q7D4M4;
DT 02-NOV-2016, integrated into UniProtKB/Swiss-Prot.
DT 01-JUL-1997, sequence version 1.
DT 03-AUG-2022, entry version 126.
DE RecName: Full=Outer membrane channel protein CpnT {ECO:0000305};
DE AltName: Full=Channel protein with necrosis-inducing toxin {ECO:0000303|PubMed:24753609};
DE Contains:
DE RecName: Full=N-terminal channel domain {ECO:0000305};
DE Contains:
DE RecName: Full=Tuberculosis necrotizing toxin {ECO:0000303|PubMed:26237511};
DE Short=TNT {ECO:0000303|PubMed:26237511};
DE AltName: Full=NAD(+) glycohydrolase {ECO:0000303|PubMed:26237511};
DE EC=3.2.2.5 {ECO:0000269|PubMed:26237511};
GN Name=cpnT {ECO:0000303|PubMed:24753609};
GN OrderedLocusNames=Rv3903c {ECO:0000312|EMBL:CCP46732.1};
OS Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv).
OC Bacteria; Actinobacteria; Corynebacteriales; Mycobacteriaceae;
OC Mycobacterium; Mycobacterium tuberculosis complex.
OX NCBI_TaxID=83332;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=9634230; DOI=10.1038/31159;
RA Cole S.T., Brosch R., Parkhill J., Garnier T., Churcher C.M., Harris D.E.,
RA Gordon S.V., Eiglmeier K., Gas S., Barry C.E. III, Tekaia F., Badcock K.,
RA Basham D., Brown D., Chillingworth T., Connor R., Davies R.M., Devlin K.,
RA Feltwell T., Gentles S., Hamlin N., Holroyd S., Hornsby T., Jagels K.,
RA Krogh A., McLean J., Moule S., Murphy L.D., Oliver S., Osborne J.,
RA Quail M.A., Rajandream M.A., Rogers J., Rutter S., Seeger K., Skelton S.,
RA Squares S., Squares R., Sulston J.E., Taylor K., Whitehead S.,
RA Barrell B.G.;
RT "Deciphering the biology of Mycobacterium tuberculosis from the complete
RT genome sequence.";
RL Nature 393:537-544(1998).
RN [2]
RP IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=21969609; DOI=10.1074/mcp.m111.011627;
RA Kelkar D.S., Kumar D., Kumar P., Balakrishnan L., Muthusamy B., Yadav A.K.,
RA Shrivastava P., Marimuthu A., Anand S., Sundaram H., Kingsbury R.,
RA Harsha H.C., Nair B., Prasad T.S., Chauhan D.S., Katoch K., Katoch V.M.,
RA Kumar P., Chaerkady R., Ramachandran S., Dash D., Pandey A.;
RT "Proteogenomic analysis of Mycobacterium tuberculosis by high resolution
RT mass spectrometry.";
RL Mol. Cell. Proteomics 10:M111.011627-M111.011627(2011).
RN [3]
RP FUNCTION, SUBUNIT, SUBCELLULAR LOCATION, PROTEOLYTIC CLEAVAGE, DISRUPTION
RP PHENOTYPE, AND MUTAGENESIS OF GLY-818.
RX PubMed=24753609; DOI=10.1073/pnas.1400136111;
RA Danilchanka O., Sun J., Pavlenok M., Maueroeder C., Speer A., Siroy A.,
RA Marrero J., Trujillo C., Mayhew D.L., Doornbos K.S., Munoz L.E.,
RA Herrmann M., Ehrt S., Berens C., Niederweis M.;
RT "An outer membrane channel protein of Mycobacterium tuberculosis with
RT exotoxin activity.";
RL Proc. Natl. Acad. Sci. U.S.A. 111:6750-6755(2014).
RN [4]
RP DISRUPTION PHENOTYPE.
RC STRAIN=H37Rv;
RX PubMed=25645841; DOI=10.1128/aac.04222-14;
RA Danilchanka O., Pires D., Anes E., Niederweis M.;
RT "The Mycobacterium tuberculosis outer membrane channel protein CpnT confers
RT susceptibility to toxic molecules.";
RL Antimicrob. Agents Chemother. 59:2328-2336(2015).
RN [5] {ECO:0007744|PDB:4QLP}
RP X-RAY CRYSTALLOGRAPHY (1.10 ANGSTROMS) OF 651-846 IN COMPLEX WITH IFT,
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, INTERACTION WITH IFT, SUBCELLULAR LOCATION, DISRUPTION
RP PHENOTYPE, AND MUTAGENESIS OF TYR-765; HIS-792; GLY-818 AND GLN-822.
RX PubMed=26237511; DOI=10.1038/nsmb.3064;
RA Sun J., Siroy A., Lokareddy R.K., Speer A., Doornbos K.S., Cingolani G.,
RA Niederweis M.;
RT "The tuberculosis necrotizing toxin kills macrophages by hydrolyzing NAD.";
RL Nat. Struct. Mol. Biol. 22:672-678(2015).
CC -!- FUNCTION: Has a dual function in uptake of nutrients and induction of
CC host cell death. The N-terminal domain (NTD) forms an outer membrane
CC channel and is used for uptake of nutrients across the outer membrane.
CC The secreted C-terminal toxic domain (TNT) acts as a glycohydrolase,
CC which hydrolyzes the essential cellular coenzyme NAD(+) in the cytosol
CC of infected macrophages, leading to necrotic host cell death. Both
CC functions are required for survival, replication and cytotoxicity of
CC M.tuberculosis in macrophages. {ECO:0000269|PubMed:24753609,
CC ECO:0000269|PubMed:26237511}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + NAD(+) = ADP-D-ribose + H(+) + nicotinamide;
CC Xref=Rhea:RHEA:16301, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:17154, ChEBI:CHEBI:57540, ChEBI:CHEBI:57967; EC=3.2.2.5;
CC Evidence={ECO:0000269|PubMed:26237511};
CC -!- ACTIVITY REGULATION: Glycohydrolase activity is completely inhibited by
CC interaction with the immunity factor for TNT (IFT). This inhibition
CC protects M.tuberculosis from self-poisoning.
CC {ECO:0000269|PubMed:26237511}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=614 uM for NAD(+) {ECO:0000269|PubMed:26237511};
CC Note=kcat is 52 sec(-1). {ECO:0000269|PubMed:26237511};
CC pH dependence:
CC Optimum pH is 6.5. {ECO:0000269|PubMed:26237511};
CC -!- SUBUNIT: Interacts with the immunity factor for TNT (IFT)
CC (PubMed:26237511). Oligomer formation is required for channel activity
CC (PubMed:24753609). {ECO:0000269|PubMed:24753609,
CC ECO:0000269|PubMed:26237511}.
CC -!- INTERACTION:
CC O05442; O05443: Rv3902c; NbExp=5; IntAct=EBI-16167127, EBI-16167151;
CC -!- SUBCELLULAR LOCATION: [N-terminal channel domain]: Cell outer membrane
CC {ECO:0000269|PubMed:24753609}.
CC -!- SUBCELLULAR LOCATION: [Tuberculosis necrotizing toxin]: Secreted
CC {ECO:0000269|PubMed:24753609}. Cell surface
CC {ECO:0000269|PubMed:24753609}. Host cytoplasm, host cytosol
CC {ECO:0000269|PubMed:26237511}. Note=Secreted into the cytosol of
CC infected macrophages while the bacteria are still confined to the
CC phagosome. Access to the macrophage cytosol depends on the ESX-1 / type
CC VII secretion system (T7SS). {ECO:0000269|PubMed:26237511}.
CC -!- PTM: The C-terminal toxic domain is cleaved, probably after integration
CC of CpnT into the outer membrane. {ECO:0000269|PubMed:24753609}.
CC -!- DISRUPTION PHENOTYPE: Deletion reduces growth on glycerol and glucose
CC as sole carbon sources. Deletion mutant does not replicate in
CC differentiated THP-1 macrophages and lacks cytotoxicity
CC (PubMed:24753609). Lack of cpnT does not increase drug resistance in
CC vitro (PubMed:25645841). Deletion mutant does not decrease NAD(+)
CC levels in infected macrophages (PubMed:26237511).
CC {ECO:0000269|PubMed:24753609, ECO:0000269|PubMed:25645841,
CC ECO:0000269|PubMed:26237511}.
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DR EMBL; AL123456; CCP46732.1; -; Genomic_DNA.
DR RefSeq; NP_218420.1; NC_000962.3.
DR RefSeq; WP_003899759.1; NZ_NVQJ01000005.1.
DR PDB; 4QLP; X-ray; 1.10 A; B=651-846.
DR PDBsum; 4QLP; -.
DR AlphaFoldDB; O05442; -.
DR SMR; O05442; -.
DR IntAct; O05442; 1.
DR STRING; 83332.Rv3903c; -.
DR PaxDb; O05442; -.
DR GeneID; 886229; -.
DR KEGG; mtu:Rv3903c; -.
DR PATRIC; fig|83332.111.peg.4347; -.
DR TubercuList; Rv3903c; -.
DR eggNOG; COG0803; Bacteria.
DR OMA; RNGLCSI; -.
DR Reactome; R-HSA-9637698; Phagocyte cell death caused by cytosolic Mtb.
DR Proteomes; UP000001584; Chromosome.
DR GO; GO:0009279; C:cell outer membrane; IEA:UniProtKB-SubCell.
DR GO; GO:0009986; C:cell surface; IEA:UniProtKB-SubCell.
DR GO; GO:0005829; C:cytosol; TAS:Reactome.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0044164; C:host cell cytosol; IEA:UniProtKB-SubCell.
DR GO; GO:0009274; C:peptidoglycan-based cell wall; HDA:UniProtKB.
DR GO; GO:0046930; C:pore complex; IEA:UniProtKB-KW.
DR GO; GO:0061810; F:NAD glycohydrolase activity; TAS:Reactome.
DR GO; GO:0015288; F:porin activity; IEA:UniProtKB-KW.
DR GO; GO:0090729; F:toxin activity; IEA:UniProtKB-KW.
DR GO; GO:0006811; P:ion transport; IEA:UniProtKB-KW.
DR GO; GO:0001907; P:killing by symbiont of host cells; TAS:Reactome.
DR InterPro; IPR025331; TNT.
DR Pfam; PF14021; TNT; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cell outer membrane; Host cytoplasm; Hydrolase;
KW Ion transport; Membrane; NAD; Porin; Reference proteome; Secreted; Toxin;
KW Transmembrane; Transmembrane beta strand; Transport; Virulence.
FT CHAIN 1..846
FT /note="Outer membrane channel protein CpnT"
FT /id="PRO_0000437782"
FT CHAIN 1..?
FT /note="N-terminal channel domain"
FT /evidence="ECO:0000305"
FT /id="PRO_0000437783"
FT CHAIN ?..846
FT /note="Tuberculosis necrotizing toxin"
FT /evidence="ECO:0000305"
FT /id="PRO_0000437784"
FT DOMAIN 751..846
FT /note="TNT"
FT /evidence="ECO:0000255"
FT REGION 1..443
FT /note="NTD"
FT /evidence="ECO:0000305|PubMed:24753609"
FT REGION 442..630
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 449..508
FT /note="Pro residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 757
FT /evidence="ECO:0000250|UniProtKB:Q4WL81"
FT ACT_SITE 822
FT /evidence="ECO:0000250|UniProtKB:Q4WL81"
FT BINDING 780
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250|UniProtKB:Q4WL81"
FT MUTAGEN 765
FT /note="Y->A: Decreases glycohydrolase activity and
FT cytotoxicity in macrophages."
FT /evidence="ECO:0000269|PubMed:26237511"
FT MUTAGEN 792
FT /note="H->N: Lack of glycohydrolase activity and of
FT cytotoxicity; when associated with K-822."
FT /evidence="ECO:0000269|PubMed:26237511"
FT MUTAGEN 818
FT /note="G->V: Unfolded. Lack of glycohydrolase activity.
FT Abolishes toxicity."
FT /evidence="ECO:0000269|PubMed:24753609,
FT ECO:0000269|PubMed:26237511"
FT MUTAGEN 822
FT /note="Q->A: 2-fold decrease in glycohydrolase activity.
FT Intermediate cytotoxicity."
FT /evidence="ECO:0000269|PubMed:26237511"
FT MUTAGEN 822
FT /note="Q->K: Lack of glycohydrolase activity and of
FT cytotoxicity; when associated with N-792."
FT /evidence="ECO:0000269|PubMed:26237511"
FT TURN 678..680
FT /evidence="ECO:0007829|PDB:4QLP"
FT HELIX 683..686
FT /evidence="ECO:0007829|PDB:4QLP"
FT HELIX 707..714
FT /evidence="ECO:0007829|PDB:4QLP"
FT STRAND 715..717
FT /evidence="ECO:0007829|PDB:4QLP"
FT STRAND 723..725
FT /evidence="ECO:0007829|PDB:4QLP"
FT HELIX 730..732
FT /evidence="ECO:0007829|PDB:4QLP"
FT STRAND 739..743
FT /evidence="ECO:0007829|PDB:4QLP"
FT HELIX 744..751
FT /evidence="ECO:0007829|PDB:4QLP"
FT STRAND 753..759
FT /evidence="ECO:0007829|PDB:4QLP"
FT STRAND 766..768
FT /evidence="ECO:0007829|PDB:4QLP"
FT HELIX 770..772
FT /evidence="ECO:0007829|PDB:4QLP"
FT HELIX 778..780
FT /evidence="ECO:0007829|PDB:4QLP"
FT HELIX 784..788
FT /evidence="ECO:0007829|PDB:4QLP"
FT STRAND 791..796
FT /evidence="ECO:0007829|PDB:4QLP"
FT STRAND 804..810
FT /evidence="ECO:0007829|PDB:4QLP"
FT STRAND 821..826
FT /evidence="ECO:0007829|PDB:4QLP"
FT HELIX 835..840
FT /evidence="ECO:0007829|PDB:4QLP"
FT STRAND 843..845
FT /evidence="ECO:0007829|PDB:4QLP"
SQ SEQUENCE 846 AA; 88335 MW; CB7DF1D77C7C9A25 CRC64;
MAPLAVDPAA LDSAGGAVVA AGAGLGAVIS SLTAALAGCA GMAGDDPAGA VFGRSYDGSA
AALVQAMSVA RNGLCNLGDG VRMSAHNYSL AEAMSDVAGR AAPLPAPPPS GCVGVGAPPS
AVGGGGGAPK GWGWVAPYIG MIWPNGDSTK LRAAAVAWRS AGTQFALTEI QSTAGPMGVI
RAQQLPEAGL IESAFADAYA STTAVVGQCH QLAAQLDAYA ARIDAVHAAV LDLLARICDP
LTGIKEVWEF LTDQDEDEIQ RIAHDIAVVV DQFSGEVDAL AAEITAVVSH AEAVITAMAD
HAGKQWDRFL HSNPVGVVID GTGQQLKGFG EEAFGMAKDS WDLGPLRASI DPFGWYRSWE
EMLTGMAPLA GLGGENAPGV VESWKQFGKS LIHWDEWTTN PNEALGKTVF DAATLALPGG
PLSKLGSKGR DILAGVRGLK ERLEPTTPHL EPPATPPRPG PQPPRIEPPE SGHPAPAPAA
KPAPVPANGP LPHSPTESKP PPVDRPAEPV APSSASAGQP RVSAATTPGT HVPHGLPQPG
EHVPAQAPPA TTLLGGPPVE SAPATAHQPQ WATTPAAPAA APHSTPGGVH STESGPHGRS
LSAHGSEPTH DGASHGSGHG SGSEPPGLHA PHREQQLAMH SNEPAGEGWH RLSDEAVDPQ
YGEPLSRHWD FTDNPADRSR INPVVAQLME DPNAPFGRDP QGQPYTQERY QERFNSVGPW
GQQYSNFPPN NGAVPGTRIA YTNLEKFLSD YGPQLDRIGG DQGKYLAIME HGRPASWEQR
ALHVTSLRDP YHAYTIDWLP EGWFIEVSEV APGCGQPGGS IQVRIFDHQN EMRKVEELIR
RGVLRQ
