Human connexin43 gap junction channels

Regulation of unitary conductances by phosphorylation

A. P. Moreno, J. C. Sáez, G. I. Fishman, David C. Spray

Research output: Contribution to journalArticle

203 Citations (Scopus)

Abstract

Connexin43 is the major gap protein in the heart and cardiovascular system. Single channel recordings of human connexin43 gap junction channels exogenously expressed in transfected SKHep1 cells demonstrate two discrete classes of channel events, with unitary conductances of predominantly 60 to 70 and 90 to 100 pS when recorded with an internal solution containing CsCl as the major current-carrying ionic species and at moderate transjunctional voltages (<60 mV). Human connexin43 expressed in SKHep1 cells displays multiple electrophoretic mobilities (apparent M(r), ≃41 to 45 kD) when resolved in Western blots. Treatment of connexin43 from these cells with alkaline phosphatase collapses the bands into a single 41-kD species; application of alkaline phosphatase to the cell interior through patch pipettes yields channels that are predominantly of the larger unitary conductance. The smaller 60- to 70-pS unitary conductance values correspond to the most common channel size seen in cultured rat cardiac myocytes; these channels were more frequently observed after treatment with the phosphatase inhibitor okadaic acid, which was shown to increase phosphorylation of human connexin43 in these cells under similar conditions. Exposure to the protein kinase inhibitor staurosporine shifted the proportion of events toward the largest unitary conductance and resulted in decreased phosphorylation of human connexin43 in seryl residues in these cells. Thus, the unitary conductance of human connexin43 gap junction channels covaries with the phosphorylation state of the protein. This change in unitary conductance appears to be a unique effect of phosphorylation on gap junction channels, since it has not been observed for other ion channels that have thus far been evaluated.

Original languageEnglish (US)
Pages (from-to)1050-1057
Number of pages8
JournalCirculation Research
Volume74
Issue number6
StatePublished - Jun 1994

Fingerprint

Connexin 43
Gap Junctions
Phosphorylation
Alkaline Phosphatase
Okadaic Acid
Staurosporine
Protein Kinase Inhibitors
Cardiovascular System
Ion Channels
Phosphoric Monoester Hydrolases
Cardiac Myocytes
Proteins
Western Blotting

Keywords

  • cDNA transfection
  • heart connexin
  • single channel conductance
  • SKHep1 cells

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Human connexin43 gap junction channels : Regulation of unitary conductances by phosphorylation. / Moreno, A. P.; Sáez, J. C.; Fishman, G. I.; Spray, David C.

In: Circulation Research, Vol. 74, No. 6, 06.1994, p. 1050-1057.

Research output: Contribution to journalArticle

Moreno, A. P. ; Sáez, J. C. ; Fishman, G. I. ; Spray, David C. / Human connexin43 gap junction channels : Regulation of unitary conductances by phosphorylation. In: Circulation Research. 1994 ; Vol. 74, No. 6. pp. 1050-1057.
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