O17J_CONGE
ID O17J_CONGE Reviewed; 82 AA.
AC X5IWS1;
DT 15-FEB-2017, integrated into UniProtKB/Swiss-Prot.
DT 15-FEB-2017, sequence version 2.
DT 25-MAY-2022, entry version 32.
DE RecName: Full=Mu-conotoxin GVIIJ {ECO:0000303|PubMed:26817840};
DE AltName: Full=Conotoxin muO-GVIIJ {ECO:0000303|PubMed:26817840};
DE AltName: Full=MuO'section sign'-GVIIJ {ECO:0000303|PubMed:26817840};
DE Flags: Precursor;
OS Conus geographus (Geography cone) (Nubecula geographus).
OC Eukaryota; Metazoa; Spiralia; Lophotrochozoa; Mollusca; Gastropoda;
OC Caenogastropoda; Neogastropoda; Conoidea; Conidae; Conus; Gastridium.
OX NCBI_TaxID=6491;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA] OF 1-75, PROTEIN SEQUENCE OF 48-59 AND 65-74,
RP MASS SPECTROMETRY, AND SUBCELLULAR LOCATION.
RC TISSUE=Venom, and Venom duct;
RX PubMed=24662800; DOI=10.1038/ncomms4521;
RA Dutertre S., Jin A.-H., Vetter I., Hamilton B., Sunagar K., Lavergne V.,
RA Dutertre V., Fry B.G., Antunes A., Venter D.J., Alewood P.F., Lewis R.J.;
RT "Evolution of separate predation- and defence-evoked venoms in carnivorous
RT cone snails.";
RL Nat. Commun. 5:3521-3521(2014).
RN [2]
RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 48-82, FUNCTION, SYNTHESIS
RP OF 48-82, DISULFIDE BOND, BROMINATION AT TRP-49, HYDROXYLATION AT PRO-53,
RP S-CYSTEINYLATION AT CYS-71, AND MASS SPECTROMETRY.
RC TISSUE=Venom, and Venom duct;
RX PubMed=24497506; DOI=10.1073/pnas.1324189111;
RA Gajewiak J., Azam L., Imperial J., Walewska A., Green B.R.,
RA Bandyopadhyay P.K., Raghuraman S., Ueberheide B., Bern M., Zhou H.M.,
RA Minassian N.A., Hagan R.H., Flinspach M., Liu Y., Bulaj G., Wickenden A.D.,
RA Olivera B.M., Yoshikami D., Zhang M.M.;
RT "A disulfide tether stabilizes the block of sodium channels by the
RT conotoxin muO[section sign]-GVIIJ.";
RL Proc. Natl. Acad. Sci. U.S.A. 111:2758-2763(2014).
RN [3]
RP SYNTHESIS OF 48-82 (WITHOUT BROMINATION).
RX PubMed=26039939; DOI=10.1021/acs.biochem.5b00390;
RA Zhang M.M., Gajewiak J., Azam L., Bulaj G., Olivera B.M., Yoshikami D.;
RT "Probing the redox states of sodium channel cysteines at the binding site
RT of muO[section sign]-conotoxin GVIIJ.";
RL Biochemistry 54:3911-3920(2015).
RN [4]
RP FUNCTION, AND SYNTHESIS OF 48-82.
RX PubMed=25632083; DOI=10.1152/jn.01004.2014;
RA Wilson M.J., Zhang M.M., Gajewiak J., Azam L., Rivier J.E., Olivera B.M.,
RA Yoshikami D.;
RT "Alpha- and beta-subunit composition of voltage-gated sodium channels
RT investigated with mu-conotoxins and the recently discovered muO'section
RT sign'-conotoxin GVIIJ.";
RL J. Neurophysiol. 113:2289-2301(2015).
RN [5]
RP STRUCTURE BY NMR OF 48-82 OF MUTANT CYS-71, SYNTHESIS OF 48-82 (WITHOUT
RP BROMINATION), FUNCTION, DISULFIDE BONDS, AND MUTAGENESIS OF TRP-49; TRP-49;
RP ASP-52; PRO-53; THR-56; LYS-59; LEU-60; ARG-61; LEU-62; TYR-63; SER-66;
RP PHE-68; ASP-70; TYR-72; THR-73; LYS-74; THR-75; LYS-77; ASP-78 AND LYS-79.
RX PubMed=26817840; DOI=10.1074/jbc.m115.697672;
RA Green B.R., Gajewiak J., Chhabra S., Skalicky J.J., Zhang M., Rivier J.E.,
RA Bulaj G., Olivera B.M., Yoshikami D., Norton R.S.;
RT "Structural basis for the inhibition of voltage-gated sodium channels by
RT conotoxin muO-GVIIJ.";
RL J. Biol. Chem. 291:7205-7220(2016).
CC -!- FUNCTION: Mu-conotoxins block voltage-gated sodium channels (Nav). This
CC toxin (GVIIJ(SSG)) blocks Nav1.1/SCN1A (Kd=11 nM), Nav1.2/SCN2A (Kd=11
CC nM), Nav1.3/SCN3A (Kd=15 nM), Nav1.4/SCN4A (Kd=4.7 nM), Nav1.6/SCN8A
CC (Kd=360 nM) and Nav1.7/SCN9A (Kd=41 nM) (PubMed:24497506,
CC PubMed:26039939). It binds the channel at the newly described site 8,
CC which is composed by two surfaces whose one contains a non-disulfide-
CC bonded cysteine (which is free to covalently bind the toxin Cys-71)
CC (PubMed:24497506). It is noteworthy that coexpression of subunits beta-
CC 2 or beta-4 (but not beta-1 or beta-3) protects rNav1.1-1.7 against
CC block by the toxin, since these subunits (thanks to their extracellular
CC domain) covalently bind to the key cysteine of the channel, thus
CC preventing the covalent binding of the toxin (PubMed:24497506,
CC PubMed:25632083). {ECO:0000269|PubMed:24497506,
CC ECO:0000269|PubMed:25632083, ECO:0000269|PubMed:26039939,
CC ECO:0000269|PubMed:26817840}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:24662800}.
CC -!- TISSUE SPECIFICITY: Expressed by the venom duct.
CC {ECO:0000305|PubMed:24662800}.
CC -!- DOMAIN: The presence of a 'disulfide through disulfide knot'
CC structurally defines this protein as a knottin.
CC {ECO:0000269|PubMed:26817840}.
CC -!- DOMAIN: The cysteine framework is VI/VII (C-C-CC-C-C). {ECO:0000305}.
CC -!- PTM: Cys-71 is a key residue that tethers to the channel by covalent
CC attachment, leading to nearly irreversible inhibition (k(off) very low)
CC (PubMed:24497506, PubMed:26817840). In order to determine the solution
CC structure without dimerization, this residue was mutated to Cys.
CC {ECO:0000269|PubMed:24497506, ECO:0000269|PubMed:26817840}.
CC -!- MASS SPECTROMETRY: Mass=3934.49; Method=MALDI;
CC Evidence={ECO:0000269|PubMed:24497506};
CC -!- MISCELLANEOUS: This toxin shows a very low affinity to Nav1.5/SCN5A
CC (Kd=207 uM) and does not show activity on rNav1.8/SCN10A.
CC {ECO:0000269|PubMed:24497506}.
CC -!- SIMILARITY: Belongs to the conotoxin O1 superfamily. {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=BAO65619.1; Type=Erroneous termination; Note=Truncated C-terminus.; Evidence={ECO:0000305};
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DR EMBL; AB910851; BAO65619.1; ALT_SEQ; mRNA.
DR PDB; 2N8H; NMR; -; A=48-82.
DR PDBsum; 2N8H; -.
DR AlphaFoldDB; X5IWS1; -.
DR BMRB; X5IWS1; -.
DR SMR; X5IWS1; -.
DR TCDB; 8.B.4.1.9; the conotoxin t (conotoxin t) family.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0008200; F:ion channel inhibitor activity; IEA:InterPro.
DR GO; GO:0017080; F:sodium channel regulator activity; IEA:UniProtKB-KW.
DR GO; GO:0090729; F:toxin activity; IEA:UniProtKB-KW.
DR InterPro; IPR004214; Conotoxin.
DR Pfam; PF02950; Conotoxin; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Bromination; Direct protein sequencing; Disulfide bond;
KW Hydroxylation; Ion channel impairing toxin; Knottin; Secreted; Signal;
KW Toxin; Voltage-gated sodium channel impairing toxin.
FT SIGNAL 1..22
FT /evidence="ECO:0000255"
FT PROPEP 23..47
FT /evidence="ECO:0000269|PubMed:24497506"
FT /id="PRO_0000438880"
FT CHAIN 48..82
FT /note="Mu-conotoxin GVIIJ"
FT /evidence="ECO:0000269|PubMed:24497506"
FT /id="PRO_5004957961"
FT SITE 59
FT /note="Functionally important residue, that binds to the
FT site 8 of the channel"
FT /evidence="ECO:0000305|PubMed:26817840"
FT SITE 61
FT /note="Functionally important residue, that binds to the
FT site 8 of the channel"
FT /evidence="ECO:0000305|PubMed:26817840"
FT SITE 63
FT /note="Functionally important residue, that binds to the
FT site 8 of the channel"
FT /evidence="ECO:0000305|PubMed:26817840"
FT SITE 71
FT /note="Functionally important residue, that binds to the
FT site 8 of the channel (distinct surface that K-59; R-61 and
FT Y-63)"
FT /evidence="ECO:0000305|PubMed:26817840"
FT MOD_RES 49
FT /note="6'-bromotryptophan"
FT /evidence="ECO:0000269|PubMed:24497506"
FT MOD_RES 53
FT /note="4-hydroxyproline"
FT /evidence="ECO:0000269|PubMed:24497506"
FT MOD_RES 71
FT /note="S-cysteinyl cysteine"
FT /evidence="ECO:0000269|PubMed:24497506"
FT DISULFID 50..65
FT /evidence="ECO:0000269|PubMed:24497506,
FT ECO:0000269|PubMed:26817840, ECO:0007744|PDB:2N8H"
FT DISULFID 57..69
FT /evidence="ECO:0000269|PubMed:24497506,
FT ECO:0000269|PubMed:26817840, ECO:0007744|PDB:2N8H"
FT DISULFID 64..76
FT /evidence="ECO:0000269|PubMed:24497506,
FT ECO:0000269|PubMed:26817840, ECO:0007744|PDB:2N8H"
FT MUTAGEN 49
FT /note="W->A: 10-fold decrease of affinity to Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 52
FT /note="D->K: No change in affinity to Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 53
FT /note="P->A: No change in affinity to Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 56
FT /note="T->A: No change in affinity to Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 59
FT /note="K->D: 280-fold decrease of affinity to
FT Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 60
FT /note="L->A: Small decrease in affinity to Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 61
FT /note="R->D: 133-fold decrease of affinity to
FT Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 62
FT /note="L->A: No change in affinity to Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 63
FT /note="Y->A: 53-fold decrease of affinity to Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 66
FT /note="S->A: Small decrease in affinity to Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 68
FT /note="F->A: No change in affinity to Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 70
FT /note="D->N: No change in affinity to Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 72
FT /note="Y->A,D,R: No change or small decrease in affinity to
FT Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 73
FT /note="T->A: No change in affinity to Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 74
FT /note="K->D: 13-fold decrease of affinity to Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 74
FT /note="K->G,F: No change in affinity to Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 75
FT /note="T->A: Small decrease in affinity to Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 77
FT /note="K->A: No change in affinity to Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 78
FT /note="D->K: No change in affinity to Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT MUTAGEN 79
FT /note="K->D: Small decrease in affinity to Nav1.2/SCN2A."
FT /evidence="ECO:0000269|PubMed:26817840"
FT STRAND 58..61
FT /evidence="ECO:0007829|PDB:2N8H"
FT STRAND 64..67
FT /evidence="ECO:0007829|PDB:2N8H"
FT TURN 71..74
FT /evidence="ECO:0007829|PDB:2N8H"
SQ SEQUENCE 82 AA; 8871 MW; D8A907002C69A971 CRC64;
MKLTCVVIVA ALLLTACQLI TALDCGGTQK HRALRSTIKL SLLRQHRGWC GDPGATCGKL
RLYCCSGFCD CYTKTCKDKS SA
