RegIIA (P00032) Protein Card

General Information

Name	RegIIA
Alternative name(s)	Reg2a
Organism	Conus regius
Organism region	Eastern Pacific
Organism diet	vermivorous
Protein Type	Wild type
Protein precursor	RegIIA precursor (5529)
Notes	Zheng et al. (2022) indicates that H14 is essential to the secondary structure of RegIIA and it plays an important role in the inhibitory activity of the peptide at human α3β4 nAChRs. Oligosaccharide chains affect the binding of RegIIA at the hα3β4 nAChR through unstable hydrogen bond interactions with H14 and by affecting the C-loop conformation of the binding sites.

Classification

Conopeptide class	conotoxin
Gene superfamily	A superfamily
Cysteine framework	I
Pharmacological family	alpha conotoxin

Sequence

GCCSHPACNVNNPHIC(nh2)

Modified residues

position	symbol	name
17	nh2	C-term amidation

Sequence evidence

protein level

Average Mass

1663.88

Monoisotopic Mass

1662.63

Isoelectric Point

8.42

Extinction Coefficient [280nm]

Activity

IC50: Nicotinic acetylcholine receptors

Target	Organism		IC50		n_hill	Agonist	Ref
α3β2	H. sapiens	(?3[P198Q]r?2)	31.0 nM	[25-38.4]	1.8	ACh (50 uM)	Kompella,S.N. et al. (2015)
			704.1 nM	[502.9-985.9]	1.2	ACh (50 uM)	Kompella,S.N. et al. (2015)
			132.4nM	[109.8-159.7]	1.5	50uM ACh	Cuny,H. et al. (2016)
	R. norvegicus		34.5 nM	[26.2-45.4]	-0.998	ACh (100 uM)	Wang,S. et al. (2022)
		(?3[E187D]?2)	7.3 nM	[5-10.8]	1.7	ACh (50 uM)	Kompella,S.N. et al. (2015)
		(?3[Q198P]?2)	520.9 nM	[400.2-678.1]	0.8	ACh (50 uM)	Kompella,S.N. et al. (2015)
		(?3[Q73G]+[V75A]?2)	29.6 nM	[23.9-36.7]	2.3	ACh (50 uM)	Kompella,S.N. et al. (2015)
		(?3?2[D73E])	37.6 nM	[29.4-48.1]	1.4	ACh (50 uM)	Kompella,S.N. et al. (2015)
		(?3?2[D165E])	32.2 nM	[26.5-39]	1.7	ACh (50 uM)	Kompella,S.N. et al. (2015)
		(?3[D97V]?2)	28.3 nM	[23.8-33.7]	1.7	ACh (50 uM)	Kompella,S.N. et al. (2015)
		(?3?2[I181V])	39.2 nM	[31.3-49]	1.1	ACh (50 uM)	Kompella,S.N. et al. (2015)
			33nM	+/-4		30uM ACh	Franco,A. et al. (2012)
			10.7 nM	[8.8-12.9]	-1.1	ACh (50 uM)	Kompella,S.N. et al. (2015)
α3β4	H. sapiens		45.6nM	[31.5-65.9]	0.9	300uM Ach	Cuny,H. et al. (2016)
			52.1 nM	[42.9-63.3]	1.9	ACh (50 uM)	Kompella,S.N. et al. (2015)
	R. norvegicus		47.3 nM	[39.5-56.6]	-1.5	ACh (50 uM)	Kompella,S.N. et al. (2015)
			97nM	+/-20		30uM ACh	Franco,A. et al. (2012)
α4β2	R. norvegicus		>1uM			30uM ACh	Franco,A. et al. (2012)
α6/α3β4β3	R. norvegicus		147 nM	[118.7-182.1]	-1.4	ACh (50 uM)	Kompella,S.N. et al. (2015)
α7	H. sapiens		210 nM	[170-270]	1.2	200 uM Ach	Yu et al. (2018)
			103nM	+/-23		200uM ACh	Franco,A. et al. (2012)
	R. norvegicus		41 nM	[33-50]	1.1	200 uM Ach	Yu et al. (2018)
			61.2 nM	[47.8-78.4]	-1.2	ACh (200 uM)	Kompella,S.N. et al. (2015)
α9α10	H. sapiens	(human a9 rat a10)	>1uM			30uM ACh	Franco,A. et al. (2012)

Synthetic variants

RegIIA[H14A]	GCCSHPACNVNNPAIC(nh2)
RegIIA[H14D]	GCCSHPACNVNNPDIC(nh2)
RegIIA[H14N]	GCCSHPACNVNNPNIC(nh2)
RegIIA[H14S]	GCCSHPACNVNNPSIC(nh2)
RegIIA[H14W]	GCCSHPACNVNNPWIC(nh2)
RegIIA[H14Y]	GCCSHPACNVNNPYIC(nh2)
RegIIA[H5D]	GCCSDPACNVNNPHIC(nh2)
RegIIA[I15A]	GCCSHPACNVNNPHAC(nh2)
RegIIA[N11A,N12A]	GCCSHPACNVAAPHIC(nh2)
RegIIA[N11A]	GCCSHPACNVANPHIC(nh2)
RegIIA[N11H]	GCCSHPACNVHNPHIC(nh2)
RegIIA[N11R]	GCCSHPACNVRNPHIC(nh2)
RegIIA[N11Y]	GCCSHPACNVYNPHIC(nh2)
RegIIA[N12A]	GCCSHPACNVNAPHIC(nh2)
RegIIA[N9A]	GCCSHPACAVNNPHIC(nh2)
RegIIA[N9F]	GCCSHPACFVNNPHIC(nh2)
RegIIA[N9R]	GCCSHPACRVNNPHIC(nh2)
RegIIA[N9W]	GCCSHPACWVNNPHIC(nh2)
RegIIA[N9Y]	GCCSHPACYVNNPHIC(nh2)
RegIIA[P13A]	GCCSHPACNVNNAHIC(nh2)
RegIIA[V10A]	GCCSHPACNANNPHIC(nh2)
RegIIA[V10F]	GCCSHPACNFNNPHIC(nh2)
RegIIA[V10Y]	GCCSHPACNYNNPHIC(nh2)

References

Franco,A., Pisarewicz,K., Moller,C., Mora,D., Fields,G.B. and Mari,F. (2006) Hyperhydroxylation: a new strategy for neuronal targeting by venomous marine molluscs Prog. Mol. Subcell. Biol. 43:83

Cuny,H., Kompella,S.N., Tae,H.S., Yu,R. and Adams,D.J. (2016) Key Structural Determinants in the Agonist Binding Loops of Human β2 and β4 Nicotinic Acetylcholine Receptor Subunits Contribute to α3β4 Subtype Selectivity of α-Conotoxins. J. Biol. Chem. 291:23779-23792

Franco,A., Kompella,S.N., Akondi,K.B., Melaun,C., Daly,N.L., Luetje,C.W., Alewood,P.F., Craik,D.J., Adams,D.J. and Marí,F. (2012) RegIIA: an α4/7-conotoxin from the venom of Conus regius that potently blocks α3β4 nAChRs. Biochem. Pharmacol. 83:419-426

Yu, J., Zhu, X., Zhang, L., Kudryavtsev, D., Kasheverov, I., Lei, Y., Zhangsun, D., Tsetlin, V. and Luo, S. (2018) Species specificity of rat and human α7 nicotinic acetylcholine receptors towards different classes of peptide and protein antagonists Neuropharmacology 139:226-237

Kompella,S.N., Hung,A., Clark,R.J., Marí,F. and Adams,D.J. (2015) Alanine scan of α-conotoxin RegIIA reveals a selective α3β4 nicotinic acetylcholine receptor antagonist J. Biol. Chem. 290:1039-1048

Kompella,S.N., Cuny,H., Hung,A. and Adams,D.J. (2015) Molecular Basis for Differential Sensitivity of α-Conotoxin RegIIA at Rat and Human Neuronal Nicotinic Acetylcholine Receptors Mol. Pharmacol. 88:993-1001

Xu,Q., Tae,H.S., Wang,Z., Jiang,T., Adams,D.J. and Yu,R. (2020) Rational Design of α-Conotoxin RegIIA Analogues Specifically Inhibiting the Human α3β2 Nicotinic Acetylcholine Receptor through Computational Scanning ACS Chem Neurosci

Zheng,M., Tae,H.S., Xue,L., Jian,T., Yu,R. (2022) Mechanism of interactions between α-conotoxin RegIIA and carbohydrates at the human α3β4 nicotinic acetylcholine receptor. Mar Life Sci Technol 4:98-105

Wang,S., Bartels,P., Zhao,C., Yousuf,A., Liu,Z., Yu,S., Bony,A.R., Ma,X., Dai,Q., Sun,T., Liu,N., Yang,M., Yu,R., Du,W., Adams,D.J., Dai,Q. (2022) A 4/8 Subtype α-Conotoxin Vt1.27 Inhibits N-Type Calcium Channels With Potent Anti-Allodynic Effect Frontiers in Pharmacology 13

Internal links

Protein Precursor	RegIIA precursor (5529)
Nucleic acids
Structure	RegIIA NMR solution structure

External links

UniProtKB/Swiss-Prot	P85013
Ncbi	P85013

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