Proteolytic processing of the L-type Ca2+ channel alpha1 1.2 subunit in neurons [version 2; referees: 3 approved]

Olivia R. Buonarati, Peter B. Henderson, Geoffrey G. Murphy, Mary C Horne, Johannes W Hell

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Background: The L-type Ca2+ channel Cav1.2 is a prominent regulator of neuronal excitability, synaptic plasticity, and gene expression. The central element of Cav1.2 is the pore-forming α11.2 subunit. It exists in two major size forms, whose molecular masses have proven difficult to precisely determine. Recent work suggests that α11.2 is proteolytically cleaved between the second and third of its four pore-forming domains (Michailidis et al,. 2014). Methods: To better determine the apparent molecular masses (MR)of the α1 1.2 size forms, extensive systematic immunoblotting of brain tissue as well as full length and C-terminally truncated α11.2 expressed in HEK293 cells was conducted using six different region–specific antibodies against α11.2. Results: The full length form of α11.2 migrated, as expected, with an apparent MR of ~250 kDa. A shorter form of comparable prevalence with an apparent MR of ~210 kDa could only be detected in immunoblots probed with antibodies recognizing α11.2 at an epitope 400 or more residues upstream of the C-terminus. Conclusions: The main two size forms of α11.2 are the full length form and a shorter form, which lacks ~350 distal C-terminal residues. Midchannel cleavage as suggested by Michailidis et al. (2014) is at best minimal in brain tissue.

Original languageEnglish (US)
Article number1166
JournalF1000Research
Volume6
DOIs
StatePublished - Jan 1 2018

Fingerprint

Molecular mass
Neurons
Neuronal Plasticity
Antibodies
HEK293 Cells
Brain
Processing
Immunoblotting
Epitopes
Tissue
Gene Expression
Gene expression
Plasticity

Keywords

  • Calpain cleavage
  • Cav1.2
  • Neuronal calcium

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

Proteolytic processing of the L-type Ca2+ channel alpha1 1.2 subunit in neurons [version 2; referees : 3 approved]. / Buonarati, Olivia R.; Henderson, Peter B.; Murphy, Geoffrey G.; Horne, Mary C; Hell, Johannes W.

In: F1000Research, Vol. 6, 1166, 01.01.2018.

Research output: Contribution to journalArticle

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