Characterization of the high-affinity verapamil binding site in a plant plasma membrane Ca2+-selective channel

M. Piñeros*, Mark Tester

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

27 Scopus citations

Abstract

Despite biochemical evidence for the existence of high-affinity phenylalkylamine receptors in higher plants, their effects on channel activity have only been demonstrated at relatively high concentrations. We have performed a quantitative single-channel analysis of the changes induced by extracellular verapamil in the rca channel [a wheat root plasma membrane Ca2+-selective channel (Pineros and Tester, 1995. Planta 195:478-488)], Concentrations as low as 0.5 μM verapamil induced a blockade of the inward current, with no evident reduction of the single-channel current amplitude. Blockade by verapamil was concentration and voltage dependent. Preliminary analysis suggested the blockade was due to a reduction in the maximum open state probability rather than a change in V0.5. Further analysis of the association and dissociation rate constants revealed a binding site located 56 to 59% down the voltage drop from the extracellular face of the channel, with a K(d)(0) of 24 to 26 μM. This results in a K(d) at -100 mV of 2 μM. Methoxyverapamil had qualitatively the same effects. This intra-pore binding site can be accessed directly from the extracellular side of the I rca channel, but apparently not from the cytosolic side.

Original languageEnglish (US)
Pages (from-to)139-145
Number of pages7
JournalJournal of Membrane Biology
Volume157
Issue number2
DOIs
StatePublished - Jun 19 1997

Keywords

  • Calcium channel
  • Methoxyverapamil
  • Verapamil
  • Wheat roots

ASJC Scopus subject areas

  • Biophysics
  • Physiology
  • Cell Biology

Fingerprint

Dive into the research topics of 'Characterization of the high-affinity verapamil binding site in a plant plasma membrane Ca<sup>2+</sup>-selective channel'. Together they form a unique fingerprint.

Cite this