A kinetic analysis of protein transport through the anthraxtoxin channel

Daniel Basilio, Paul K. Kienker, Stephen W. Briggs, Alan Finkelstein

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Anthrax toxin is composed of three proteins: a translocase heptameric channel, (PA63)7, formed from protective antigen (PA), which allows the other two proteins, lethal factor (LF) and edema factor (EF), to translocate across a host cell's endosomal membrane, disrupting cellular homeostasis. (PA63)7 incorporated into planar phospholipid bilayer membranes forms a channel capable of transporting LF and EF. Protein translocation through the channel can be driven by voltage on a timescale of seconds. A characteristic of the translocation of LFN, the N-terminal 263 residues of LF, is its S-shaped kinetics. Because all of the translocation experiments reported in the literature have been performed with more than one LFN molecule bound to most of the channels, it is not clear whether the S-shaped kinetics are an intrinsic characteristic of translocation kinetics or are merely a consequence of the translocation in tandem of two or three LFNs. In this paper, we show both in macroscopic and single-channel experiments that even with only one LFN bound to the channel, the translocation kinetics are S shaped. As expected, the translocation rate is slower with more than one LFN bound. We also present a simple electrodiffusion model of translocation in which LFN is represented as a charged rod that moves subject to both Brownian motion and an applied electric field. The cumulative distribution of first-passage times of the rod past the end of the channel displays S-shaped kinetics with a voltage dependence in agreement with experimental data.

Original languageEnglish (US)
Pages (from-to)521-531
Number of pages11
JournalJournal of General Physiology
Volume137
Issue number6
DOIs
StatePublished - Jun 2011

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Protein Transport
Phospholipids
Proteins
Homeostasis
Cell Membrane
Antigens
Membranes
edema factor

ASJC Scopus subject areas

  • Physiology

Cite this

A kinetic analysis of protein transport through the anthraxtoxin channel. / Basilio, Daniel; Kienker, Paul K.; Briggs, Stephen W.; Finkelstein, Alan.

In: Journal of General Physiology, Vol. 137, No. 6, 06.2011, p. 521-531.

Research output: Contribution to journalArticle

Basilio, D, Kienker, PK, Briggs, SW & Finkelstein, A 2011, 'A kinetic analysis of protein transport through the anthraxtoxin channel', Journal of General Physiology, vol. 137, no. 6, pp. 521-531. https://doi.org/10.1085/jgp.201110627
Basilio, Daniel ; Kienker, Paul K. ; Briggs, Stephen W. ; Finkelstein, Alan. / A kinetic analysis of protein transport through the anthraxtoxin channel. In: Journal of General Physiology. 2011 ; Vol. 137, No. 6. pp. 521-531.
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