TX15_GRAPO
ID TX15_GRAPO Reviewed; 34 AA.
AC P0DL72;
DT 25-OCT-2017, integrated into UniProtKB/Swiss-Prot.
DT 25-OCT-2017, sequence version 1.
DT 25-MAY-2022, entry version 17.
DE RecName: Full=Toxin GTx1-15 {ECO:0000250|UniProtKB:P0DJA9};
DE AltName: Full=Beta/omega-theraphotoxin-Gr2a {ECO:0000305};
DE Short=Beta/omega-TRTX-Gr2a {ECO:0000305};
DE AltName: Full=Toxin GpTx-1 {ECO:0000303|PubMed:25658507};
OS Grammostola porteri (Tarantula spider) (Lasiodora porteri).
OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Chelicerata; Arachnida; Araneae;
OC Mygalomorphae; Theraphosidae; Grammostola.
OX NCBI_TaxID=1749325;
RN [1]
RP PROTEIN SEQUENCE, FUNCTION, SYNTHESIS, STRUCTURE BY NMR, MUTAGENESIS OF
RP ASP-1; LEU-3; GLY-4; PHE-5; MET-6; ARG-7; LYS-8; ILE-10; PRO-11; ASP-12;
RP ASN-13; LYS-15; ARG-18; PRO-19; ASN-20; LEU-21; VAL-22; SER-24; ARG-25;
RP THR-26; HIS-27; LYS-28; TRP-29; LYS-31; TYR-32; VAL-33 AND PHE-34,
RP AMIDATION AT PHE-34, AND MASS SPECTROMETRY.
RC TISSUE=Venom;
RX PubMed=25658507; DOI=10.1021/jm501765v;
RA Murray J.K., Ligutti J., Liu D., Zou A., Poppe L., Li H., Andrews K.L.,
RA Moyer B.D., McDonough S.I., Favreau P., Stoecklin R., Miranda L.P.;
RT "Engineering potent and selective analogues of GpTx-1, a tarantula venom
RT peptide antagonist of the Na(V)1.7 sodium channel.";
RL J. Med. Chem. 58:2299-2314(2015).
RN [2]
RP FUNCTION.
RX PubMed=26999206; DOI=10.3390/toxins8030078;
RA Deuis J.R., Wingerd J.S., Winter Z., Durek T., Dekan Z., Sousa S.R.,
RA Zimmermann K., Hoffmann T., Weidner C., Nassar M.A., Alewood P.F.,
RA Lewis R.J., Vetter I.;
RT "Analgesic effects of GpTx-1, PF-04856264 and CNV1014802 in a mouse model
RT of Nav1.7-mediated pain.";
RL Toxins 8:1-19(2016).
CC -!- FUNCTION: Potent voltage-gated sodium channel blocker
CC (PubMed:25658507). Potently inhibits the voltage-gated sodium channels
CC Nav1.7/SCN9A (IC(50)=0.58-10 nM) (PubMed:25658507, PubMed:26999206).
CC Also shows a moderate activity on Nav1.1/SCN1A (IC(50)=6 nM),
CC Nav1.2/SCN2A (IC(50)=5-128 nM), Nav1.3/SCN3A (IC(50)=20.3-170 nM), and
CC Nav1.6/SCN8A (IC(50)=17-20.1 nM) (PubMed:25658507, PubMed:26999206).
CC Shows an unclear inhibition of Nav1.4/SCN4A (IC(50)=200 nM to >10 uM),
CC Nav1.5/SCN5A (IC(50)=140 nM to >10 uM) and Nav1.8/SCN10A (IC(50)=68-
CC 12200 nM) (PubMed:25658507, PubMed:26999206). Weakly blocks the low
CC voltage-gated calcium channels Cav3.1/CACNA1G (30% inhibition of the
CC peak current at 9.8 nM) (By similarity). shows moderate affinity for
CC lipid bilayers (By similarity). In vivo, when tested on the OD1-induced
CC mouse model of Nav1.7/SCN9A-mediated pain, the toxin is effective when
CC co-administered with OD1, but lacks efficacy when delivered
CC systemically (PubMed:26999206). {ECO:0000250|UniProtKB:P0DJA9,
CC ECO:0000250|UniProtKB:P0DL73, ECO:0000269|PubMed:25658507,
CC ECO:0000269|PubMed:26999206}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:25658507}.
CC -!- TISSUE SPECIFICITY: Expressed by the venom gland.
CC {ECO:0000305|PubMed:25658507}.
CC -!- DOMAIN: The presence of a 'disulfide through disulfide knot'
CC structurally defines this protein as a knottin.
CC {ECO:0000269|PubMed:25658507}.
CC -!- DOMAIN: This toxin is amphipathic in nature with a hydrophobic face on
CC one side of the molecule (composed of residues 5-Phe-Met-6 and 27-His--
CC Phe-34) and a hydrophilic (mostly cationic) face on the opposite side
CC (composed of residues 10-Ile--Lys-15 and 18-Arg--Pro-19).
CC {ECO:0000269|PubMed:25658507}.
CC -!- MASS SPECTROMETRY: Mass=4073.9; Method=MALDI; Note=Average mass.;
CC Evidence={ECO:0000269|PubMed:25658507};
CC -!- MISCELLANEOUS: The mutant [Phe5Ala]GpTx1 may be used as a tool for
CC probing Nav1.7/SCN9A inhibition in vivo. {ECO:0000269|PubMed:25658507}.
CC -!- MISCELLANEOUS: The primary structure of the mature peptide is identical
CC to that of Gtx1-15 from Grammostola rosea (AC P0DJA9) and Gtx1-15 from
CC Paraphysa scrofa (AC P0DL73). {ECO:0000305}.
CC -!- SIMILARITY: Belongs to the neurotoxin 10 (Hwtx-1) family. 08 (Gtx1-15)
CC subfamily. {ECO:0000305}.
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DR PDB; 6MK5; NMR; -; A=1-34.
DR PDBsum; 6MK5; -.
DR AlphaFoldDB; P0DL72; -.
DR BMRB; P0DL72; -.
DR SMR; P0DL72; -.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0005246; F:calcium channel regulator activity; IEA:UniProtKB-KW.
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; IPR011696; Huwentoxin-1.
DR InterPro; IPR013140; Huwentoxin_CS1.
DR Pfam; PF07740; Toxin_12; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Amidation; Calcium channel impairing toxin;
KW Direct protein sequencing; Disulfide bond; Ion channel impairing toxin;
KW Knottin; Secreted; Toxin; Voltage-gated calcium channel impairing toxin;
KW Voltage-gated sodium channel impairing toxin.
FT CHAIN 1..34
FT /note="Toxin GTx1-15"
FT /evidence="ECO:0000269|PubMed:25658507"
FT /id="PRO_0000441929"
FT SITE 5
FT /note="Important for activity and sodium channel
FT selectivity"
FT /evidence="ECO:0000305|PubMed:25658507"
FT SITE 6
FT /note="Important for activity"
FT /evidence="ECO:0000305|PubMed:25658507"
FT SITE 26
FT /note="Important for activity and sodium channel
FT selectivity"
FT /evidence="ECO:0000305|PubMed:25658507"
FT SITE 27
FT /note="Important for activity"
FT /evidence="ECO:0000305|PubMed:25658507"
FT SITE 28
FT /note="Important for activity and sodium channel
FT selectivity"
FT /evidence="ECO:0000305|PubMed:25658507"
FT SITE 29
FT /note="Important for activity"
FT /evidence="ECO:0000305|PubMed:25658507"
FT SITE 31
FT /note="Important for activity"
FT /evidence="ECO:0000305|PubMed:25658507"
FT SITE 32
FT /note="Important for activity"
FT /evidence="ECO:0000305|PubMed:25658507"
FT SITE 34
FT /note="Important for activity"
FT /evidence="ECO:0000305|PubMed:25658507"
FT MOD_RES 34
FT /note="Phenylalanine amide"
FT /evidence="ECO:0000269|PubMed:25658507"
FT DISULFID 2..17
FT /evidence="ECO:0000269|PubMed:25658507"
FT DISULFID 9..23
FT /evidence="ECO:0000269|PubMed:25658507"
FT DISULFID 16..30
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 1
FT /note="D->A: Decrease in selectivity for hNav1.7/SCN9A over
FT hNav1.4/SCN4A. No change in activity on hNav1.7/SCN9A and
FT hNav1.5/SCN5A and 1.5-fold increase in activity on
FT hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 1
FT /note="D->AD: 4-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and loss of activity on
FT hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 3
FT /note="L->A: 5-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and loss of activity on
FT hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 4
FT /note="G->A: 3-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and 1.2-fold decrease in
FT activity on hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 5..6
FT /note="FM->AF: 1200-fold increase in selectivity for
FT hNav1.7/SCN9A over hNav1.4/SCN4A; compound 71."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 5
FT /note="F->A: 2.5-fold increase in selectivity for
FT hNav1.7/SCN9A over hNav1.4/SCN4A. ~5-fold decrease in
FT activity on hNav1.7/SCN9A, more than 5-fold decrease in
FT activity on hNav1.5/SCN5A and ~25-fold decrease in activity
FT on hNav1.4/SCN4A (depending on the method used). 3.6-fold
FT decrease in activity on TTX-S sodium channels from mouse
FT DRG neurons."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 6
FT /note="M->A: 5-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and 1.4-fold decrease in
FT activity on hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 7
FT /note="R->A: 10-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and loss of activity on
FT hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 8
FT /note="K->A: 5-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and loss of activity on
FT hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 10
FT /note="I->A: 2-fold decrease in activity on hNav1.7/SCN9A,
FT and no change on hNav1.5/SCN5A and hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 11
FT /note="P->A: No significant change on hNav1.7/SCN9A,
FT hNav1.5/SCN5A and hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 12
FT /note="D->A: No change in activity on hNav1.7/SCN9A and
FT hNav1.5/SCN5A and 3-fold increase in activity on
FT hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 13
FT /note="N->A: 2-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 15
FT /note="K->A: 3-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and 1.7-fold decrease in
FT activity on hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 18
FT /note="R->A: No change in activity on hNav1.7/SCN9A and
FT hNav1.4/SCN4A, and decrease in activity on hNav1.5/SCN5A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 19
FT /note="P->A: No change in activity on hNav1.7/SCN9A,
FT decrease in activity on hNav1.5/SCN5A, and 1.8-fold
FT increase in activity on hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 20
FT /note="N->A: 8-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and loss of activity on
FT hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 21
FT /note="L->A: Decrease in activity on hNav1.7/SCN9A,
FT hNav1.4/SCN4A, and hNav1.5/SCN5A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 22
FT /note="V->A: 4-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and 1.4-fold decrease in
FT activity on hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 24
FT /note="S->A: 5-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and 1.4-fold increase in
FT activity on hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 25
FT /note="R->A: 4-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and loss of activity on
FT hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 26
FT /note="T->A: 3-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and 1.2-fold decrease in
FT activity on hNav1.4/SCN4A. 1200-fold increase in
FT selectivity for hNav1.7/SCN9A over hNav1.4/SCN4A; compound
FT 71."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 27
FT /note="H->A: 11-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and 1.5-fold decrease in
FT activity on hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 28
FT /note="K->A: 5-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and loss of activity on
FT hNav1.4/SCN4A. 1200-fold increase in selectivity for
FT hNav1.7/SCN9A over hNav1.4/SCN4A; compound 71."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 29
FT /note="W->A: Loss of activity on hNav1.7/SCN9A and
FT hNav1.4/SCN4A, and no change on hNav1.5/SCN5A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 31
FT /note="K->A: Loss of activity on hNav1.7/SCN9A and
FT hNav1.4/SCN4A, and no change on hNav1.5/SCN5A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 32
FT /note="Y->A: 9-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and loss of activity on
FT hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 33
FT /note="V->A: 2-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and 1.7-fold decrease in
FT activity on hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 34
FT /note="F->A: 13-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and loss of activity on
FT hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT MUTAGEN 34
FT /note="F->FA: 5-fold decrease in activity on hNav1.7/SCN9A,
FT no change on hNav1.5/SCN5A and loss of activity on
FT hNav1.4/SCN4A."
FT /evidence="ECO:0000269|PubMed:25658507"
FT HELIX 11..13
FT /evidence="ECO:0007829|PDB:6MK5"
FT TURN 18..20
FT /evidence="ECO:0007829|PDB:6MK5"
FT STRAND 21..24
FT /evidence="ECO:0007829|PDB:6MK5"
FT TURN 25..28
FT /evidence="ECO:0007829|PDB:6MK5"
FT STRAND 29..32
FT /evidence="ECO:0007829|PDB:6MK5"
SQ SEQUENCE 34 AA; 4081 MW; DDEF29032F4A84E2 CRC64;
DCLGFMRKCI PDNDKCCRPN LVCSRTHKWC KYVF
