The number of subunits comprising the channel formed by the T domain of diphtheria toxin.
In the presence of a low pH environment, the channel-forming T domain of diphtheria toxin undergoes a conformational change that allows for both its own insertion into planar lipid bilayers and the translocation of the toxin's catalytic domain across them. Given that the T domain contributes only three transmembrane segments, and the channel is permeable to ions as large as glucosamine(+) and NAD(-), it would appear that the channel must be a multimer. Yet, there is substantial circumstantial evidence that the channel may be formed from a single subunit. To test the hypothesis that the channel formed by the T domain of diphtheria toxin is monomeric, we made mixtures of two T domain constructs whose voltage-gating characteristics differ, and then observed the gating behavior of the mixture's single channels in planar lipid bilayers. One of these constructs contained an NH(2)-terminal hexahistidine (H6) tag that blocks the channel at negative voltages; the other contained a COOH-terminal H6 tag that blocks the channel at positive voltages. If the channel is constructed from multiple T domain subunits, one expects to see a population of single channels from this mixture that are blocked at both positive and negative voltages. The observed single channels were blocked at either negative or positive voltages, but never both. Therefore, we conclude that the T domain channel is monomeric.
Gordon
M
M
Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Finkelstein
A
A
D000818:Animals; D002462:Cell Membrane / Q000502:physiology; D017931:DNA Primers; D004167:Diphtheria Toxin / Q000737:chemistry* / Q000494:pharmacology*; D004594:Electrophysiology; D066257:Heparin-binding EGF-like Growth Factor; D006863:Hydrogen-Ion Concentration; D036341:Intercellular Signaling Peptides and Proteins; D015640:Ion Channel Gating; D008051:Lipid Bilayers; D008563:Membrane Lipids / Q000502:physiology; D010539:Permeability; D010957:Plasmids; D011956:Receptors, Cell Surface / Q000502:physiology*
D017931:DNA Primers; D004167:Diphtheria Toxin; D066257:Heparin-binding EGF-like Growth Factor; D036341:Intercellular Signaling Peptides and Proteins; D008051:Lipid Bilayers; D008563:Membrane Lipids; D011956:Receptors, Cell Surface