title: Peptide- and proton-driven allosteric clamps catalyze anthrax toxin translocation across membranes.
abstract: Anthrax toxin is an intracellularly acting toxin in which sufficient information is available regarding the structure of its transmembrane channel, allowing for detailed investigation of models of translocation. Anthrax toxin, comprising three proteins-protective antigen (PA), lethal factor (LF), and edema factor-translocates large proteins across membranes. Here we show that the PA translocase channel has a transport function in which its catalytic active sites operate allosterically. We find that the phenylalanine clamp (ϕ-clamp), the known conductance bottleneck in the PA translocase, gates as either a more closed state or a more dilated state. Thermodynamically, the two channel states have >300-fold different binding affinities for an LF-derived peptide. The change in clamp thermodynamics requires distant α-clamp and ϕ-clamp sites. Clamp allostery and translocation are more optimal for LF peptides with uniform stereochemistry, where the least allosteric and least efficiently translocated peptide had a mixed stereochemistry. Overall, the kinetic results are in less agreement with an extended-chain Brownian ratchet model but, instead, are more consistent with an allosteric helix-compression model that is dependent also on substrate peptide coil-to-helix/helix-to-coil cooperativity.
authors: Department of Microbial Pathogenesis, School of Dentistry, University of Maryland, Baltimore, MD 21201.
authors: Debasis
authors: D
authors: Das
authors: Department of Microbial Pathogenesis, School of Dentistry, University of Maryland, Baltimore, MD 21201 bkrantz@umaryland.edu.
authors: Bryan A
authors: BA
authors: Krantz
mesh_terms: D000494:Allosteric Regulation; D000942:Antigens, Bacterial / Q000378:metabolism*; D001427:Bacterial Toxins / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism*; D010649:Phenylalanine / Q000378:metabolism; D010713:Phosphatidylcholines / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D017433:Protein Structure, Secondary; D021381:Protein Transport; D011522:Protons; D013237:Stereoisomerism; D013816:Thermodynamics
chemical_list: D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D011522:Protons; C030325:anthrax toxin; D010649:Phenylalanine
keywords: Brownian ratchet; allostery; anthrax toxin; helix-compression model; planar bilayer electrophysiology