CA1A_CONRE
ID CA1A_CONRE Reviewed; 32 AA.
AC P0C1D0; Q1WJB2;
DT 16-MAY-2006, integrated into UniProtKB/Swiss-Prot.
DT 16-MAY-2006, sequence version 1.
DT 25-MAY-2022, entry version 54.
DE RecName: Full=Alpha-conotoxin RgIA {ECO:0000303|PubMed:16445293, ECO:0000303|PubMed:18242183};
DE Flags: Precursor; Fragment;
OS Conus regius (Crown cone).
OC Eukaryota; Metazoa; Spiralia; Lophotrochozoa; Mollusca; Gastropoda;
OC Caenogastropoda; Neogastropoda; Conoidea; Conidae; Conus; Stephanoconus.
OX NCBI_TaxID=101314;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], SYNTHESIS OF 20-32, AND FUNCTION.
RC TISSUE=Hepatopancreas;
RX PubMed=16445293; DOI=10.1021/bi0520129;
RA Ellison M., Haberlandt C., Gomez-Casati M.E., Watkins M., Elgoyhen A.B.,
RA McIntosh J.M., Olivera B.M.;
RT "Alpha-RgIA: a novel conotoxin that specifically and potently blocks the
RT alpha9alpha10 nAChR.";
RL Biochemistry 45:1511-1517(2006).
RN [2]
RP FUNCTION, AND BIOASSAY.
RX PubMed=17101979; DOI=10.1073/pnas.0608715103;
RA Vincler M., Wittenauer S., Parker R., Ellison M., Olivera B.M.,
RA McIntosh J.M.;
RT "Molecular mechanism for analgesia involving specific antagonism of
RT alpha9alpha10 nicotinic acetylcholine receptors.";
RL Proc. Natl. Acad. Sci. U.S.A. 103:17880-17884(2006).
RN [3]
RP FUNCTION ON GABA(B) RECEPTOR, AND SYNTHESIS OF 20-32.
RX PubMed=18945902; DOI=10.1523/jneurosci.3594-08.2008;
RA Callaghan B., Haythornthwaite A., Berecki G., Clark R.J., Craik D.J.,
RA Adams D.J.;
RT "Analgesic alpha-conotoxins Vc1.1 and Rg1A inhibit N-type calcium channels
RT in rat sensory neurons via GABAB receptor activation.";
RL J. Neurosci. 28:10943-10951(2008).
RN [4]
RP FUNCTION, MUTAGENESIS OF ARG-32, SYNTHESIS OF 20-32, AND SYNTHESIS OF
RP CYCLIC ANALOGS.
RX PubMed=21888386; DOI=10.1021/jm201060r;
RA Halai R., Callaghan B., Daly N.L., Clark R.J., Adams D.J., Craik D.J.;
RT "Effects of cyclization on stability, structure, and activity of alpha-
RT conotoxin RgIA at the alpha9alpha10 nicotinic acetylcholine receptor and
RT GABA(B) receptor.";
RL J. Med. Chem. 54:6984-6992(2011).
RN [5]
RP FUNCTION, AND SYNTHESIS OF 20-32.
RX PubMed=22774872; DOI=10.1111/j.1471-4159.2012.07867.x;
RA Azam L., McIntosh J.M.;
RT "Molecular basis for the differential sensitivity of rat and human
RT alpha9alpha10 nAChRs to alpha-conotoxin RgIA.";
RL J. Neurochem. 122:1137-1144(2012).
RN [6]
RP BIOASSAY, AND SYNTHESIS OF 20-32.
RX PubMed=25008370; DOI=10.1016/j.pain.2014.06.023;
RA Di Cesare Mannelli L., Cinci L., Micheli L., Zanardelli M., Pacini A.,
RA McIntosh J.M., Ghelardini C.;
RT "Alpha-conotoxin RgIA protects against the development of nerve injury-
RT induced chronic pain and prevents both neuronal and glial derangement.";
RL Pain 155:1986-1995(2014).
RN [7]
RP FUNCTION, SITES ARG-26 AND ARG-30, 3D-STRUCTURE MODELING, AND SYNTHESIS OF
RP 20-32.
RX PubMed=25740413; DOI=10.1124/mol.114.096511;
RA Azam L., Papakyriakou A., Zouridakis M., Giastas P., Tzartos S.J.,
RA McIntosh J.M.;
RT "Molecular interaction of alpha-conotoxin RgIA with the rat alpha9alpha10
RT nicotinic acetylcholine receptor.";
RL Mol. Pharmacol. 87:855-864(2015).
RN [8]
RP ERRATUM OF PUBMED:25740413.
RX PubMed=27559150; DOI=10.1124/mol.114.096511err;
RA Azam L., Papakyriakou A., Zouridakis M., Giastas P., Tzartos S.J.,
RA McIntosh J.M.;
RT "Corrections to 'Molecular interaction of alpha-conotoxin RgIA with the rat
RT alpha9alpha10 nicotinic acetylcholine receptor'.";
RL Mol. Pharmacol. 90:415-417(2016).
RN [9]
RP PHARMACEUTICAL, MUTAGENESIS OF SER-23; ARG-28; TYR-29; ARG-30 AND ARG-32,
RP AND SYNTHESIS OF 20-32.
RX PubMed=28223528; DOI=10.1073/pnas.1621433114;
RA Romero H.K., Christensen S.B., Di Cesare Mannelli L., Gajewiak J.,
RA Ramachandra R., Elmslie K.S., Vetter D.E., Ghelardini C., Iadonato S.P.,
RA Mercado J.L., Olivera B.M., McIntosh J.M.;
RT "Inhibition of alpha9alpha10 nicotinic acetylcholine receptors prevents
RT chemotherapy-induced neuropathic pain.";
RL Proc. Natl. Acad. Sci. U.S.A. 114:E1825-E1832(2017).
RN [10]
RP FUNCTION, MUTAGENESIS OF CYS-31, AND PHARMACEUTICAL.
RX PubMed=32101438; DOI=10.1021/acs.jmedchem.9b01536;
RA Liang J., Tae H.S., Xu X., Jiang T., Adams D.J., Yu R.;
RT "Dimerization of alpha-conotoxins as a strategy to enhance the inhibition
RT of the human alpha7 and alpha9alpha10 nicotinic acetylcholine Receptors.";
RL J. Med. Chem. 63:2974-2985(2020).
RN [11]
RP STRUCTURE BY NMR OF 20-32, SYNTHESIS OF 20-32, AND DISULFIDE BOND.
RX PubMed=18242183; DOI=10.1016/j.febslet.2008.01.027;
RA Clark R.J., Daly N.L., Halai R., Nevin S.T., Adams D.J., Craik D.J.;
RT "The three-dimensional structure of the analgesic alpha-conotoxin, RgIA.";
RL FEBS Lett. 582:597-602(2008).
RN [12]
RP STRUCTURE BY NMR OF 20-32, DISULFIDE BOND, MUTAGENESIS OF SER-23; ASP-24;
RP PRO-25; ARG-26; ARG-28; TYR-29 AND ARG-32, AND SYNTHESIS OF 20-32.
RX PubMed=18295795; DOI=10.1016/j.jmb.2008.01.082;
RA Ellison M., Feng Z.P., Park A.J., Zhang X., Olivera B.M., McIntosh J.M.,
RA Norton R.S.;
RT "Alpha-RgIA, a novel conotoxin that blocks the alpha9alpha10 nAChR:
RT structure and identification of key receptor-binding residues.";
RL J. Mol. Biol. 377:1216-1227(2008).
RN [13]
RP STRUCTURE BY NMR OF 20-32, SYNTHESIS OF 20-32, AND SYNTHESIS OF
RP NON-REDUCIBLE ANALOGS.
RX PubMed=25393758; DOI=10.1021/jm501126u;
RA Chhabra S., Belgi A., Bartels P., van Lierop B.J., Robinson S.D.,
RA Kompella S.N., Hung A., Callaghan B.P., Adams D.J., Robinson A.J.,
RA Norton R.S.;
RT "Dicarba analogues of alpha-conotoxin RgIA. Structure, stability, and
RT activity at potential pain targets.";
RL J. Med. Chem. 57:9933-9944(2014).
CC -!- FUNCTION: This toxin target two types of receptors, the nicotinic
CC acetylcholine receptor (nAChR) and the G-protein-coupled receptor
CC GABA(B). It specifically inhibits the alpha-9-alpha-10/CHRNA9-CHRNA10
CC nAChR, with preference for rat receptors (PubMed:16445293,
CC PubMed:21888386, PubMed:22774872, PubMed:25740413, PubMed:28223528,
CC PubMed:18242183, PubMed:18295795). It interacts with the alpha-
CC 10(+)/alpha-9(-)interface of the receptor (PubMed:25740413). It shows a
CC two order of magnitude species difference potency for the rat versus
CC human alpha-9-alpha-10 nAChR, due to the Thr-86 located in the alpha-9
CC nAChR subunit (PubMed:22774872). This toxin also shows inhibition of
CC high voltage-activated (HVA) calcium channels (Cav2.2) by acting on
CC GABA(B) receptors (GABBR1 and GABBR2) (PubMed:18945902,
CC PubMed:21888386). In vivo, this toxin produces an acute antinociceptive
CC effect in peripheral nerve-injured rats, which may be related to the
CC inhibition of immune cell buildup at the site of nerve injury
CC (PubMed:17101979). In addition, when intramuscularly injected into rats
CC following chronic constriction injury of the sciatic nerve, this toxin
CC protects peripheral nervous tissues as well as prevents central
CC maladaptive plasticity by inhibiting glial cell activation
CC (PubMed:25008370). {ECO:0000269|PubMed:16445293,
CC ECO:0000269|PubMed:17101979, ECO:0000269|PubMed:18242183,
CC ECO:0000269|PubMed:18295795, ECO:0000269|PubMed:18945902,
CC ECO:0000269|PubMed:21888386, ECO:0000269|PubMed:22774872,
CC ECO:0000269|PubMed:25008370, ECO:0000269|PubMed:25740413,
CC ECO:0000269|PubMed:28223528}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000250}.
CC -!- TISSUE SPECIFICITY: Expressed by the venom duct. {ECO:0000305}.
CC -!- DOMAIN: The cysteine framework is I (CC-C-C). Alpha4/3 pattern.
CC {ECO:0000305}.
CC -!- PTM: The disulfide bond CysI-CysIII is important for alpha-9-alpha-10
CC subtype inhibition, whereas the bond CysII-CysIV contributes to GABA(B)
CC modulation. {ECO:0000269|PubMed:25393758}.
CC -!- PHARMACEUTICAL: The derivative RgIA4 (S23T; R28Citrulline; Y29(3-I-Y);
CC R30Q; R32Y) is under preclinical studies by Kineta under the name KCP-
CC 400. It is tested to prevent chronic cancer chemotherapy-induced
CC neuropathic pain. {ECO:0000269|PubMed:28223528}.
CC -!- PHARMACEUTICAL: RgIA[Del13]dimer is an analog therapeutically
CC interesting because it is the first dual inhibitor that potently and
CC selectively inhibits alpha-9-alpha-10/CHRNA9-CHRNA10 and alpha-7/CHRNA7
CC subtypes, which are both involved in the etiology of several cancers.
CC {ECO:0000305|PubMed:32101438}.
CC -!- MISCELLANEOUS: This toxin shows a weak activity on alpha-7 nAChR
CC (IC(50)=3310-4660 nM) (PubMed:16445293, PubMed:18295795). It also shows
CC a very weak activity on alpha-2-beta-2, alpha-2-beta-4, alpha-3-beta-4,
CC alpha-3-beta-2, alpha-4-beta-2, alpha-4-beta-4 and alpha-6/alpha-3-
CC beta-2-beta-3 nAChRs (IC(50)>10 uM) (PubMed:16445293).
CC {ECO:0000269|PubMed:16445293, ECO:0000269|PubMed:18295795}.
CC -!- MISCELLANEOUS: The cis-[2,8]-dicarba analog is significantly more
CC stable and less susceptible to proteolytic degradation than native
CC RgIA. {ECO:0000269|PubMed:25393758}.
CC -!- MISCELLANEOUS: Replacement of Arg-28 by a Citrulline residue strongly
CC increases the potency on human CHRNA9-CHRNA10 nAChR. Replacement of
CC Tyr-29 by a monoido-Tyr (3-I-Y) residue very strongly increases the
CC potency on human CHRNA9-CHRNA10 nAChR. {ECO:0000269|PubMed:28223528}.
CC -!- MISCELLANEOUS: The synthetic variant RgIA4 (S23T; R28Citrulline; Y29(3-
CC I-Y); R30Q; R32Y) is very potent on human CHRNA9-CHRNA10 (IC(50)=1.5
CC nM) and the most selective among all other synthetic variants tested
CC (IC(50)>10 uM on all receptors tested, and (IC(50)=1.8 uM) on alpha-
CC 7/CHRNA9 nAChR). {ECO:0000269|PubMed:28223528}.
CC -!- SIMILARITY: Belongs to the conotoxin A superfamily. {ECO:0000305}.
CC -!- CAUTION: Has the same mature sequence than Reg1e (AC P85011). RgIA
CC could therefore be the precursor of Reg1e, except that RgIA does not
CC have the C-terminal Gly residue required for C-terminal amidation of
CC Reg1e. {ECO:0000305}.
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DR EMBL; DQ239610; ABB55879.1; -; Genomic_DNA.
DR PDB; 2JUQ; NMR; -; A=20-32.
DR PDB; 2JUR; NMR; -; A=20-32.
DR PDB; 2JUS; NMR; -; A=20-32.
DR PDB; 2JUT; NMR; -; A=20-32.
DR PDB; 2MTO; NMR; -; A=20-32.
DR PDB; 2MTT; NMR; -; A=20-30.
DR PDB; 6HY7; X-ray; 2.26 A; B=20-32.
DR PDBsum; 2JUQ; -.
DR PDBsum; 2JUR; -.
DR PDBsum; 2JUS; -.
DR PDBsum; 2JUT; -.
DR PDBsum; 2MTO; -.
DR PDBsum; 2MTT; -.
DR PDBsum; 6HY7; -.
DR AlphaFoldDB; P0C1D0; -.
DR SMR; P0C1D0; -.
DR ConoServer; 593; RgIA precursor.
DR EvolutionaryTrace; P0C1D0; -.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0035792; C:host cell postsynaptic membrane; IEA:UniProtKB-KW.
DR GO; GO:0030550; F:acetylcholine receptor inhibitor activity; IEA:UniProtKB-KW.
DR GO; GO:0099106; F:ion channel regulator activity; IEA:UniProtKB-KW.
DR GO; GO:0090729; F:toxin activity; IEA:UniProtKB-KW.
DR InterPro; IPR018072; Conotoxin_a-typ_CS.
DR PROSITE; PS60014; ALPHA_CONOTOXIN; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Acetylcholine receptor inhibiting toxin; Disulfide bond;
KW G-protein coupled receptor impairing toxin; Ion channel impairing toxin;
KW Neurotoxin; Pharmaceutical; Postsynaptic neurotoxin; Secreted; Toxin.
FT PROPEP <1..19
FT /evidence="ECO:0000250"
FT /id="PRO_0000234829"
FT PEPTIDE 20..32
FT /note="Alpha-conotoxin RgIA"
FT /evidence="ECO:0000305|PubMed:16445293,
FT ECO:0000305|PubMed:18242183, ECO:0000305|PubMed:18295795,
FT ECO:0000305|PubMed:18945902, ECO:0000305|PubMed:21888386,
FT ECO:0000305|PubMed:22774872, ECO:0000305|PubMed:25008370,
FT ECO:0000305|PubMed:25393758, ECO:0000305|PubMed:25740413"
FT /id="PRO_0000234830"
FT SITE 24
FT /note="Key residue for potent inhibition of the rat CHRNA9-
FT CHRNA10 nAChR"
FT /evidence="ECO:0000269|PubMed:18295795"
FT SITE 25
FT /note="Key residue for potent inhibition of the rat CHRNA9-
FT CHRNA10 nAChR"
FT /evidence="ECO:0000269|PubMed:18295795"
FT SITE 26
FT /note="Binds to the Pro-224 and Asp-225 of the rat nAChR
FT CHRNA10 subunit"
FT /evidence="ECO:0000305|PubMed:25740413"
FT SITE 26
FT /note="Key residue for potent inhibition of the rat CHRNA9-
FT CHRNA10 nAChR"
FT /evidence="ECO:0000269|PubMed:18295795"
FT SITE 28
FT /note="Key residue for potent inhibition of the rat CHRNA9-
FT CHRNA10 nAChR"
FT /evidence="ECO:0000269|PubMed:18295795"
FT SITE 30
FT /note="Binds to the Glu-221 of the rat nAChR CHRNA10
FT subunit"
FT /evidence="ECO:0000305|PubMed:25740413"
FT DISULFID 21..27
FT /evidence="ECO:0000269|PubMed:18242183,
FT ECO:0000269|PubMed:18295795, ECO:0000312|PDB:2JUQ,
FT ECO:0000312|PDB:2JUR, ECO:0000312|PDB:2JUS,
FT ECO:0000312|PDB:2JUT"
FT DISULFID 22..31
FT /evidence="ECO:0000269|PubMed:18242183,
FT ECO:0000269|PubMed:18295795, ECO:0000312|PDB:2JUQ,
FT ECO:0000312|PDB:2JUR, ECO:0000312|PDB:2JUS,
FT ECO:0000312|PDB:2JUT"
FT MUTAGEN 23
FT /note="S->A: 1.7-fold less potent to inhibit both rat
FT CHRNA9-CHRNA10 and CHRNA7 nAChR."
FT /evidence="ECO:0000269|PubMed:18295795"
FT MUTAGEN 23
FT /note="S->T: Extremely important increase in potency on
FT human CHRNA9-CHRNA10 nAChR."
FT /evidence="ECO:0000269|PubMed:28223528"
FT MUTAGEN 24
FT /note="D->E: 783-fold and 8.3-fold less potent to inhibit
FT rat CHRNA9-CHRNA10 and CHRNA7 nAChR, respectively."
FT /evidence="ECO:0000269|PubMed:18295795"
FT MUTAGEN 25
FT /note="P->V: 480-fold and 5.8-fold less potent to inhibit
FT rat CHRNA9-CHRNA10 and CHRNA7 nAChR, respectively."
FT /evidence="ECO:0000269|PubMed:18295795"
FT MUTAGEN 26
FT /note="R->K: 1523-fold and 14.2-fold less potent to inhibit
FT rat CHRNA9-CHRNA10 and CHRNA7 nAChR, respectively."
FT /evidence="ECO:0000269|PubMed:18295795"
FT MUTAGEN 28
FT /note="R->A: 1511-fold less potent to inhibit the rat
FT CHRNA9-CHRNA10 nAChR and 5.8-fold more potent to inhibit
FT the rat CHRNA7 nAChR."
FT /evidence="ECO:0000269|PubMed:18295795"
FT MUTAGEN 28
FT /note="R->K: No significant change in potency on human
FT CHRNA9-CHRNA10 nAChR."
FT /evidence="ECO:0000269|PubMed:28223528"
FT MUTAGEN 29
FT /note="Y->W: 1.3-fold less potent to inhibit the rat
FT CHRNA9-CHRNA10 nAChR and 1.1-fold more potent to inhibit
FT the rat CHRNA7 nAChR."
FT /evidence="ECO:0000269|PubMed:18295795"
FT MUTAGEN 29
FT /note="Y->W: Relatively important increase in potency on
FT human CHRNA9-CHRNA10 nAChR."
FT /evidence="ECO:0000269|PubMed:28223528"
FT MUTAGEN 30
FT /note="R->Q: Extremely important increase in potency on
FT human CHRNA9-CHRNA10 nAChR."
FT /evidence="ECO:0000269|PubMed:28223528"
FT MUTAGEN 31
FT /note="C->CGRRRRGGCCSDPRCRYRC: RgIA[Del13]dimer; Important
FT increase of activity and specificity on both alpha-9-alpha-
FT 10/CHRNA9-CHRNA10 and alpha-7/CHRNA7 nAChRs."
FT MUTAGEN 32
FT /note="R->GGAAGAG: With cyclization, improvement in the
FT stability, small increase in inhibition of human CHRNA9/rat
FT CHRNA10 nAChR and no change in GABA(B)/N-type calcium
FT channels (cRgIA-7)."
FT /evidence="ECO:0000269|PubMed:21888386"
FT MUTAGEN 32
FT /note="R->K: No significant change in potency on human
FT CHRNA9-CHRNA10 nAChR."
FT /evidence="ECO:0000269|PubMed:28223528"
FT MUTAGEN 32
FT /note="R->Q: No significant change in potency on human
FT CHRNA9-CHRNA10 nAChR."
FT /evidence="ECO:0000269|PubMed:28223528"
FT MUTAGEN 32
FT /note="R->Y: Extremely important increase in potency on
FT human CHRNA9-CHRNA10 nAChR."
FT /evidence="ECO:0000269|PubMed:28223528"
FT MUTAGEN 32
FT /note="Missing: RgIA# with amidated C-31; no change in
FT activity on rat CHRNA9-CHRNA10 nAChR and 2.8-fold increase
FT in potency to inhibit rat CHRNA7 nAChR. No change in
FT inhibition of human-CHRNA9/rat-CHRNA10 nAChR and increase
FT in inhibition of GABA(B)/N-type calcium channels
FT (RgIA[delR])."
FT /evidence="ECO:0000269|PubMed:18295795,
FT ECO:0000269|PubMed:21888386"
FT NON_TER 1
FT /evidence="ECO:0000305|PubMed:16445293"
FT HELIX 21..23
FT /evidence="ECO:0007829|PDB:6HY7"
FT HELIX 25..27
FT /evidence="ECO:0007829|PDB:2JUR"
FT TURN 29..31
FT /evidence="ECO:0007829|PDB:2JUQ"
SQ SEQUENCE 32 AA; 3725 MW; 4EC5B132037316F7 CRC64;
SNKRKNAAML DMIAQHAIRG CCSDPRCRYR CR
