PIIIA (P02228) Protein Card

General Information

Name	PIIIA
Alternative name(s)	P3.7,P3a
Organism	Conus purpurascens
Organism region	Eastern Pacific
Organism diet	piscivorous
Protein Type	Wild type
Protein precursor	PIIIA precursor (1407)
Notes	Bind to site 1 in the Na channel pore based on competitive binding assay using saxitoxin (Shon et al 1998). 10 μM rapidly blocks A-fiber but not C-fiber compound action potentials in mouse sciatic nerve [Wilson,M.J., Zhang,M (2015)].

Classification

Conopeptide class	conotoxin
Gene superfamily	M superfamily
Cysteine framework	III
Pharmacological family	mu conotoxin

Sequence

ZRLCCGFOKSCRSRQCKOHRCC(nh2)
Modified residues	position symbol name 1 Z Pyroglutamic acid 8 O 4-Hydroxyproline 18 O 4-Hydroxyproline 23 nh2 C-term amidation
Sequence evidence	nucleic acid level
Average Mass	NA
Monoisotopic Mass	NA
Isoelectric Point	12.99
Extinction Coefficient [280nm]	NA

Activity

IC50: Voltage-Gated Sodium Channels

Target	Organism		IC50		Agonist	Ref
Nav1.1	R. norvegicus		120nM	+/-13		Wilson,M.J. et al. (2011)
Nav1.2	R. norvegicus		690nM			Safo,P. et al. (2000)
			620nM	+/-70		Wilson,M.J. et al. (2011)
Nav1.3	R. norvegicus		3.2uM	+/-0.8		Wilson,M.J. et al. (2011)
Nav1.4	R. norvegicus	(PIIIA-3 (connectivity 1-2 3-4 5-6))	203.7nM	+/-25.7		Tietze,A.A. et al. (2012)
		(PIIIA-1 (connectivity 1-5 2-6 3-4))	46.7nM	+/-6.5		Tietze,A.A. et al. (2012)
		(PIIIA-2 (connectivity 1-4 2-5 3-6))	103.2nM	+/-9.9		Tietze,A.A. et al. (2012)
			36nM	+/-8		Wilson,M.J. et al. (2011)
			41nM			Safo,P. et al. (2000)
Nav1.5	R. norvegicus		>10uM			Wilson,M.J. et al. (2011)
Nav1.6	M. musculus		100nM	+/-20		Wilson,M.J. et al. (2011)
Nav1.7	H. sapiens		3.1uM			Safo,P. et al. (2000)
	R. norvegicus		6.2uM			Safo,P. et al. (2000)
			>10uM			Wilson,M.J. et al. (2011)
Nav1.8	R. norvegicus		>10uM			Wilson,M.J. et al. (2011)
NavBh	B. halodurans		5 pM	+/-1	0.81	Finol-Urdaneta et al. (2019)

Kd: Voltage-Gated Sodium Channels

Target	Organism		Kd		Ref
Nav1.1	R. norvegicus		53nM	+/-4	Zhang,M.M. et al. (2013)
		(+NaVβ1)	14nM	+/-2	Zhang,M.M. et al. (2013)
		(+NaVβ3)	17nM	+/-3	Zhang,M.M. et al. (2013)
		(+NaVβ4)	370nM	+/-110	Zhang,M.M. et al. (2013)
		(+NaVβ2)	140nM	+/-20	Zhang,M.M. et al. (2013)
Nav1.2	R. norvegicus	(+NaVβ3)	220nM	+/-20	Zhang,M.M. et al. (2013)
		(+NaVβ2)	650nM	+/-50	Zhang,M.M. et al. (2013)
		(+NaVβ4)	1.10uM	+/-0.05	Zhang,M.M. et al. (2013)
			620nM	+/-40	Zhang,M.M. et al. (2013)
		(+NaVβ1)	240nM	+/-30	Zhang,M.M. et al. (2013)
Nav1.6	R. norvegicus	(+NaVβ2)	243nM	+/-58	Zhang,M.M. et al. (2013)
			81nM	+/-32	Zhang,M.M. et al. (2013)
		(+NaVβ4)	951nM	+/-151	Zhang,M.M. et al. (2013)
		(+NaVβ1)	5nM	+/-1	Zhang,M.M. et al. (2013)
		(+NaVβ3)	9nM	+/-2	Zhang,M.M. et al. (2013)

Percentage inhibition: Voltage-Gated Potassium Channels

Target	Organism	% inhibition		Concentration	Ref
Kv1.1	R. norvegicus	73.0	+/-5.1	10 uM	Leipold,E. et al. (2017)
Kv1.2	R. norvegicus	0		10 uM	Leipold,E. et al. (2017)
Kv1.3	R. norvegicus	15		10 uM	Leipold,E. et al. (2017)
Kv1.4	R. norvegicus	4		10 uM	Leipold,E. et al. (2017)
Kv1.5	R. norvegicus	0		10 uM	Leipold,E. et al. (2017)
Kv1.6	R. norvegicus	80.3	+/-2.9	10 uM	Leipold,E. et al. (2017)
Kv2.1	R. norvegicus	3		10 uM	Leipold,E. et al. (2017)

Percentage inhibition: Voltage-Gated Sodium Channels

Target	Organism	% inhibition		Concentration	Ref
Nav1.2	R. norvegicus	75		10 uM	Leipold,E. et al. (2017)
Nav1.4	R. norvegicus	98		10 uM	Leipold,E. et al. (2017)
Nav1.5	R. norvegicus	15		10 uM	Leipold,E. et al. (2017)
Nav1.7	R. norvegicus	45		10 uM	Leipold,E. et al. (2017)
Nav TTX-R	R. pipiens	8	+/-10	5uM	Zhang,M.M. et al. (2006)

Synthetic variants

PIIIA [G6K]	ZRLCCKFOKSCRSRQCKOHRCC(nh2)
PIIIA [K17A]	ZRLCCAFOKSCRSRQCAOHRCC(nh2)
PIIIA [K17Q]	ZRLCCAFOKSCRSRQCQOHRCC(nh2)
PIIIA [K17R]	ZRLCCGFOKSCRSRQCROHRCC(nh2)
PIIIA [R12A]	ZRLCCAFOKSCASRQCKOHRCC(nh2)
PIIIA [R12K]	ZRLCCGFOKSCKSRQCKOHRCC(nh2)
PIIIA [R12Q]	ZRLCCAFOKSCQSRQCKOHRCC(nh2)
PIIIA [R14A]	ZRLCCGFOKSCRSAQCKOHRCC(nh2)
PIIIA [R14K]	ZRLCCGFOKSCRSKQCKOHRCC(nh2)
PIIIA [R14Q]	ZRLCCGFOKSCRSQQCKOHRCC(nh2)
PIIIA [R20A]	ZRLCCAFOKSCRSRQCKOHACC(nh2)
PIIIA [R2A]	ZALCCGFOKSCRSRQCKOHRCC(nh2)
PIIIA [S13D]	ZRLCCAFOKSCRDRQCKOHRCC(nh2)

References

Shon,K.-J., Olivera,B.M., Watkins,M., Jacobsen,R.B., Gray,W.R., Floresca,C.Z., Cruz,L.J., Hillyard,D.R., Brink,A., Terlau,H. and Yoshikami,D. (1998) mu-Conotoxin PIIIA, a new peptide for discriminating among tetrodotoxin-sensitive Na channel subtypes J. Neurosci. 18:4473-4481

Safo,P., Rosenbaum,T., Shcherbatko,A., Choi,D.Y., Han,E., Toledo-Aral,J.J., Olivera,B.M., Brehm,P. and Mandel,G. (2000) Distinction among neuronal subtypes of voltage-activated sodium channels by mu-conotoxin PIIIA J. Neurosci. 20:76-80

Nielsen,K.J., Watson,M., Adams,D.J., Hammarstroem,A.K., Gage,P.W., Hill,J.M., Craik,D.J., Thomas,L., Adams,D., Alewood,P.F. and Lewis,R.J. (2002) Solution structure of mu-conotoxin PIIIA, a preferential inhibitor of persistent tetrodotoxin-sensitive sodium channels J. Biol. Chem. 277:27247-27255

Wilson,M.J., Yoshikami,D., Azam,L., Gajewiak,J., Olivera,B.M., Bulaj,G. and Zhang,M.M. (2011) μ-Conotoxins that differentially block sodium channels NaV1.1 through 1.8 identify those responsible for action potentials in sciatic nerve. Proc. Natl. Acad. Sci. U.S.A. 108:10302-10307

Zhang,M.M., Wilson,M.J., Azam,L., Gajewiak,J., Rivier,J.E., Bulaj,G., Olivera,B.M. and Yoshikami,D. (2013) Co-expression of Na(V)β subunits alters the kinetics of inhibition of voltage-gated sodium channels by pore-blocking μ-conotoxins. Br. J. Pharmacol. 168:1597-1610

Tietze,A.A., Tietze,D., Ohlenschläger,O., Leipold,E., Ullrich,F., Kühl,T., Mischo,A., Buntkowsky,G., Görlach,M., Heinemann,S.H. and Imhof,D. (2012) Structurally diverse μ-conotoxin PIIIA isomers block sodium channel NaV 1.4. Angew. Chem. Int. Ed. Engl. 51:4058-4061

Wilson,M.J., Zhang,M.M., Gajewiak,J., Azam,L., Rivier,J.E., Olivera,B.M. and Yoshikami,D. (2015) α- and β-subunit composition of voltage-gated sodium channels investigated with μ-conotoxins and the recently discovered μO§-conotoxin GVIIJ. J Neurophysiol 113:2289-2301

Finol-Urdaneta, R.K., McArthur, J.R., Korkosh, V.S., Huang, S., McMaster, D., Glavica, R., Tikhonov, D.B., Zhorov, B.S. and French, R.J. (2019) Extremely Potent Block of Bacterial Voltage-Gated Sodium Channels by μ-Conotoxin PIIIA Marine drugs 17:510-527

Himaya, S.W.A., Mari, F. and Lewis, R.J. (2018) Accelerated proteomic visualization of individual predatory venoms of Conus purpurascens reveals separately evolved predation-evoked venom cabals Scientific reports 8:1-8

Heimer, P., Tietze, A.A., Bäuml, C.A., Resemann, A., Mayer, F.J., Suckau, D., Ohlenschläger, O., Tietze, D. and Imhof, D. (2018) Conformational μ-conotoxin PIIIA isomers revisited: Impact of cysteine pairing on disulfide-bond assignment and structure elucidation Analytical chemistry 90:3321-3327

Chen, F., Huang, W., Jiang, T. and Yu, R. (2018) Determination of the μ-Conotoxin PIIIA Specificity Against Voltage-Gated Sodium Channels from Binding Energy Calculations Marine drugs 16:153

Leipold,E., Ullrich,F., Thiele,M., Tietze,A.A., Terlau,H., Imhof,D. and Heinemann,S.H. (2017) Subtype-specific block of voltage-gated K+ channels by μ-conopeptides Biochem. Biophys. Res. Commun. 482:1135-1140

Paul George,A.A., Heimer,P., Maaß,A., Hamaekers,J., Hofmann-Apitius,M., Biswas,A. and Imhof,D. (2018) Insights into the Folding of Disulfide-Rich μ-Conotoxins ACS Omega 3:12330-12340

Heimer,P., Schmitz,T., Bäuml,C.A. and Imhof,D. (2018) Synthesis and Structure Determination of μ-Conotoxin PIIIA Isomers with Different Disulfide Connectivities. J Vis Exp

Schmitz,T., Pengelley,S., Belau,E. and Imhof,D. (2020) LC-Trapped Ion Mobility Spectrometry-TOF MS Differentiation of 2- and 3-Disulfide-Bonded Isomers of the µ-Conotoxin PIIIA. Anal Chem 92:10920-10924

Internal links

Protein Precursor	PIIIA precursor (1407)
Nucleic acids
Structure	NMR SOLUTION STRUCTURE OF PIIIA TOXIN, NMR, 20 STRUCTURES NMR solution structure of PIIIA connectivity 1-5 2-6 3-4 NMR solution structure of PIIIA connectivity 1-4 2-5 3-6 NMR solution structure of PIIIA connectivity 1-2 3-4 5-6

External links

Ncbi

1R9I_A

Tools

Please cite:
Kaas Q, Yu R, Jin AH, Dutertre S and Craik DJ. ConoServer: updated content, knowledge, and discovery tools in the conopeptide database. Nucleic Acids Research (2012) 40(Database issue):D325-30
Kaas Q, Westermann JC, Halai R, Wang CK and Craik DJ. ConoServer, a database for conopeptide sequences and structures. Bioinformatics (2008) 24(3):445-6

ConoServer is managed at the Institute of Molecular Bioscience IMB, Brisbane, Australia.

The database and computational tools found on this website may be used for academic research only, provided that it is referred to ConoServer, the database of conotoxins (http://www.conoserver.org) and the above reference is cited. For any other use please contact David Craik (d.craik@imb.uq.edu.au).

Contacts:
David Craik
Quentin Kaas

Last updated: Saturday 23 September 2023