A kinetic analysis of protein transport through the anthrax toxin channel

• Published in: The Journal of general physiology Vol. 137; no. 6; pp. 521 - 531
• Main Authors: Basilio, Daniel, Kienker, Paul K, Briggs, Stephen W, Finkelstein, Alan
• Format: Journal Article
• Language: English
• Published: United States Rockefeller University Press 01.06.2011; The Rockefeller University Press
• Subjects: Anthrax; Antigens; Antigens, Bacterial - chemistry; Antigens, Bacterial - metabolism; Bacillus anthracis - metabolism; Bacterial Toxins - chemistry; Bacterial Toxins - metabolism; Brownian motion; Cell Membrane; Cells; Kinetics; Lipids; Membranes; Models, Molecular; Mutation; Protein Conformation; Protein Transport - physiology; Proteins; Toxins
• ISSN: 0022-1295; 1540-7748
• DOI: 10.1085/jgp.201110627

Anthrax toxin is composed of three proteins: a translocase heptameric channel, (PA(63))(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. (PA(63))(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 LF(N), 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 LF(N) 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 LF(N)s. In this paper, we show both in macroscopic and single-channel experiments that even with only one LF(N) bound to the channel, the translocation kinetics are S shaped. As expected, the translocation rate is slower with more than one LF(N) bound. We also present a simple electrodiffusion model of translocation in which LF(N) 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.