Voltage-dependent block of anthrax toxin channels in planar phospholipid bilayer membranes by symmetric tetraalkylammonium ions

Effects on macroscopic conductance

Robert O. Blaustein, Alan Finkelstein

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

In a recent paper (Blaustein, R. O., T. M. Koehler, R. J. Collier, and A. Finkelstein. 1989. Proc. Natl. Acad. Sci. USA. 86:2209-2213) we described the general channel-forming properties of the PA65 fragment of anthrax toxin in planar phospholipid bilayer membranes. In the present paper we extend our previous studies of the permeability properties of this channel, using a series of symmetric tetraalkylammonium (TAA) ions. Our main finding is that at micromolar concentrations on either the cis (toxin-containing) or trans side of a membrane containing many (>1,000) channels, these ions, ranging in size from tetramethylammonium to tetrahexylammonium, induce a voltage-dependent reduction of membrane conductance. (We attribute a similar voltage-dependent reduction of membrane conductance by millimolar concentrations of HEPES to a cationic form of this buffer present at micromolar concentrations.) In going from large negative to large positive voltages (on the TAA side) one sees that the conductance first decreases from its value in the absence of TAA, reaches a minimum, and then rises back at larger positive voltages toward the level in the absence of TAA. Our interpretation of this behavior is that these symmetric TAA ions block the cation-selective PA65 channel in a voltage-dependent manner. We postulate that there is a single site within the channel to which TAA ions can bind and thereby block the passage of the major current-carrying ion (potassium). A blocking ion is driven into the site by modest positive voltages, but is driven off the site and through the channel by larger positive voltages, thus explaining the relief of block. (In the accompanying paper [Blaustein, R. O., E. J. A. Lea, and A. Finkelstein. 1990. J. Gen. Physiol. 96:921-942] we confirm this interpretation of the data by analysis at the single-channel level.) This means that these blocking ions can pass through the channel; the permeability to tetrahexylammonium, the largest ion studied, implies that the narrowest part of the channel has a diameter of at least 11 Å.

Original languageEnglish (US)
Pages (from-to)905-919
Number of pages15
JournalJournal of General Physiology
Volume96
Issue number5
StatePublished - Nov 1990

Fingerprint

Phospholipids
Ions
Membranes
Permeability
HEPES
anthrax toxin
Ion Channels
Cations
Potassium
Buffers

ASJC Scopus subject areas

  • Physiology

Cite this

Voltage-dependent block of anthrax toxin channels in planar phospholipid bilayer membranes by symmetric tetraalkylammonium ions : Effects on macroscopic conductance. / Blaustein, Robert O.; Finkelstein, Alan.

In: Journal of General Physiology, Vol. 96, No. 5, 11.1990, p. 905-919.

Research output: Contribution to journalArticle

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