Evidence for a 95 kDa short form of the α(1A) subunit associated with the ω-conotoxin MVIIC receptor of the P/Q-type Ca2+ channels

Victoria E.S. Scott, Ricardo Felix, Jyothi Arikkath, Kevin P. Campbell

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Neuronal voltage-dependent Ca2+ channels have been isolated previously and shown to contain a primary α1 pore-forming subunit as well as auxiliary α2δ and β subunits, in addition to an uncharacterized 95 kDa protein. In the present study, using multiple approaches, we have extensively characterized the molecular structure of the 95 kDa protein. Separation of the P/Q- and N-type neuronal Ca2+ channels showed that the 95 kDa protein is associated exclusively with the ω-Conotoxin MVIIC receptor of the P/Q- type channels. Analysis of purified synaptic plasma membranes and the isolated P/Q-type channels, using α(1A)-specific antibodies, suggested a structural relationship between the α(1A) subunit and the 95 kDa protein. This finding was supported by protein-protein interaction data, which revealed that the β subunit can associate with the 95 kDa protein in addition to the α(1A) subunit. Changes in electrophoretic mobility after enzymatic treatment with Endo F indicated that the 95 kDa protein is glycosylated. Furthermore, microsequencing of the 95 kDa protein yielded 13 peptide sequences, all of which are present in the first half of the α(1A) subunit up to amino acid 829 of the cytoplasmic linker between repeats II and III. Taken together, our results strongly suggest that the 95 kDa glycoprotein associated with the P/Q-type Ca2+ channels is a short form of the α(1A) subunit.

Original languageEnglish (US)
Pages (from-to)641-647
Number of pages7
JournalJournal of Neuroscience
Volume18
Issue number2
StatePublished - Jan 29 1998

Fingerprint

Conotoxins
Proteins
Synaptic Membranes
Molecular Structure
Glycoproteins
Cell Membrane

Keywords

  • 95 kDa subunit
  • N-type Ca channel
  • P/Q-type Ca channels
  • α(1A) subunit
  • β subunit
  • ω-Conotoxin GVIA
  • ω-Conotoxin MVIIC

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Evidence for a 95 kDa short form of the α(1A) subunit associated with the ω-conotoxin MVIIC receptor of the P/Q-type Ca2+ channels. / Scott, Victoria E.S.; Felix, Ricardo; Arikkath, Jyothi; Campbell, Kevin P.

In: Journal of Neuroscience, Vol. 18, No. 2, 29.01.1998, p. 641-647.

Research output: Contribution to journalArticle

@article{6a371005194c4614bb4713c703d9580e,
title = "Evidence for a 95 kDa short form of the α(1A) subunit associated with the ω-conotoxin MVIIC receptor of the P/Q-type Ca2+ channels",
abstract = "Neuronal voltage-dependent Ca2+ channels have been isolated previously and shown to contain a primary α1 pore-forming subunit as well as auxiliary α2δ and β subunits, in addition to an uncharacterized 95 kDa protein. In the present study, using multiple approaches, we have extensively characterized the molecular structure of the 95 kDa protein. Separation of the P/Q- and N-type neuronal Ca2+ channels showed that the 95 kDa protein is associated exclusively with the ω-Conotoxin MVIIC receptor of the P/Q- type channels. Analysis of purified synaptic plasma membranes and the isolated P/Q-type channels, using α(1A)-specific antibodies, suggested a structural relationship between the α(1A) subunit and the 95 kDa protein. This finding was supported by protein-protein interaction data, which revealed that the β subunit can associate with the 95 kDa protein in addition to the α(1A) subunit. Changes in electrophoretic mobility after enzymatic treatment with Endo F indicated that the 95 kDa protein is glycosylated. Furthermore, microsequencing of the 95 kDa protein yielded 13 peptide sequences, all of which are present in the first half of the α(1A) subunit up to amino acid 829 of the cytoplasmic linker between repeats II and III. Taken together, our results strongly suggest that the 95 kDa glycoprotein associated with the P/Q-type Ca2+ channels is a short form of the α(1A) subunit.",
keywords = "95 kDa subunit, N-type Ca channel, P/Q-type Ca channels, α(1A) subunit, β subunit, ω-Conotoxin GVIA, ω-Conotoxin MVIIC",
author = "Scott, {Victoria E.S.} and Ricardo Felix and Jyothi Arikkath and Campbell, {Kevin P.}",
year = "1998",
month = "1",
day = "29",
language = "English (US)",
volume = "18",
pages = "641--647",
journal = "Journal of Neuroscience",
issn = "0270-6474",
publisher = "Society for Neuroscience",
number = "2",

}

TY - JOUR

T1 - Evidence for a 95 kDa short form of the α(1A) subunit associated with the ω-conotoxin MVIIC receptor of the P/Q-type Ca2+ channels

AU - Scott, Victoria E.S.

AU - Felix, Ricardo

AU - Arikkath, Jyothi

AU - Campbell, Kevin P.

PY - 1998/1/29

Y1 - 1998/1/29

N2 - Neuronal voltage-dependent Ca2+ channels have been isolated previously and shown to contain a primary α1 pore-forming subunit as well as auxiliary α2δ and β subunits, in addition to an uncharacterized 95 kDa protein. In the present study, using multiple approaches, we have extensively characterized the molecular structure of the 95 kDa protein. Separation of the P/Q- and N-type neuronal Ca2+ channels showed that the 95 kDa protein is associated exclusively with the ω-Conotoxin MVIIC receptor of the P/Q- type channels. Analysis of purified synaptic plasma membranes and the isolated P/Q-type channels, using α(1A)-specific antibodies, suggested a structural relationship between the α(1A) subunit and the 95 kDa protein. This finding was supported by protein-protein interaction data, which revealed that the β subunit can associate with the 95 kDa protein in addition to the α(1A) subunit. Changes in electrophoretic mobility after enzymatic treatment with Endo F indicated that the 95 kDa protein is glycosylated. Furthermore, microsequencing of the 95 kDa protein yielded 13 peptide sequences, all of which are present in the first half of the α(1A) subunit up to amino acid 829 of the cytoplasmic linker between repeats II and III. Taken together, our results strongly suggest that the 95 kDa glycoprotein associated with the P/Q-type Ca2+ channels is a short form of the α(1A) subunit.

AB - Neuronal voltage-dependent Ca2+ channels have been isolated previously and shown to contain a primary α1 pore-forming subunit as well as auxiliary α2δ and β subunits, in addition to an uncharacterized 95 kDa protein. In the present study, using multiple approaches, we have extensively characterized the molecular structure of the 95 kDa protein. Separation of the P/Q- and N-type neuronal Ca2+ channels showed that the 95 kDa protein is associated exclusively with the ω-Conotoxin MVIIC receptor of the P/Q- type channels. Analysis of purified synaptic plasma membranes and the isolated P/Q-type channels, using α(1A)-specific antibodies, suggested a structural relationship between the α(1A) subunit and the 95 kDa protein. This finding was supported by protein-protein interaction data, which revealed that the β subunit can associate with the 95 kDa protein in addition to the α(1A) subunit. Changes in electrophoretic mobility after enzymatic treatment with Endo F indicated that the 95 kDa protein is glycosylated. Furthermore, microsequencing of the 95 kDa protein yielded 13 peptide sequences, all of which are present in the first half of the α(1A) subunit up to amino acid 829 of the cytoplasmic linker between repeats II and III. Taken together, our results strongly suggest that the 95 kDa glycoprotein associated with the P/Q-type Ca2+ channels is a short form of the α(1A) subunit.

KW - 95 kDa subunit

KW - N-type Ca channel

KW - P/Q-type Ca channels

KW - α(1A) subunit

KW - β subunit

KW - ω-Conotoxin GVIA

KW - ω-Conotoxin MVIIC

UR - http://www.scopus.com/inward/record.url?scp=0031972927&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031972927&partnerID=8YFLogxK

M3 - Article

C2 - 9425006

AN - SCOPUS:0031972927

VL - 18

SP - 641

EP - 647

JO - Journal of Neuroscience

JF - Journal of Neuroscience

SN - 0270-6474

IS - 2

ER -