Proteolytic processing of the L-type Ca2+ channel alpha11.2 subunit in neurons

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

Research output: Contribution to journalArticle

5 Citations (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 α 11.2 subunit. It exists in two major size forms, whose molecular masses have proven difficult to precisely determine. Recent work suggests that α 11.2 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 (M R)of the α 11.2 size forms, extensive systematic immunoblotting of brain tissue as well as full length and C-terminally truncated α 11.2 expressed in HEK293 cells was conducted using six different region-specific antibodies against α 11.2. Results: The full length form of α 11.2 migrated, as expected, with an apparent M R of ~250 kDa. A shorter form of comparable prevalence with an apparent M R of ~210 kDa could only be detected in immunoblots probed with antibodies recognizing α 11.2 at an epitope 400 or more residues upstream of the C-terminus. Conclusions: The main two size forms of α 11.2 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 - 2017

Fingerprint

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

Keywords

  • Calpain cleavage
  • Cav1.2
  • Neuronal calcium

ASJC Scopus subject areas

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

Cite this

Proteolytic processing of the L-type Ca2+ channel alpha11.2 subunit in neurons. / Hell, Johannes W; Buonarati, Olivia R.; Henderson, Peter B.; Murphy, Geoffrey G.; Horne, Mary C.

In: F1000Research, Vol. 6, 1166, 2017.

Research output: Contribution to journalArticle

Hell, Johannes W ; Buonarati, Olivia R. ; Henderson, Peter B. ; Murphy, Geoffrey G. ; Horne, Mary C. / Proteolytic processing of the L-type Ca2+ channel alpha11.2 subunit in neurons. In: F1000Research. 2017 ; Vol. 6.
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