{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,12]],"date-time":"2026-04-12T13:01:37Z","timestamp":1775998897369,"version":"3.50.1"},"reference-count":31,"publisher":"Rockefeller University Press","issue":"6","content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2011,6,1]]},"abstract":"<jats:p>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\u2019s 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.<\/jats:p>","DOI":"10.1085\/jgp.201110627","type":"journal-article","created":{"date-parts":[[2011,5,30]],"date-time":"2011-05-30T13:28:49Z","timestamp":1306762129000},"page":"521-531","update-policy":"https:\/\/doi.org\/10.1085\/jgp.crossmarkpolicy","source":"Crossref","is-referenced-by-count":22,"title":["A kinetic analysis of protein transport through the anthrax toxin channel"],"prefix":"10.1085","volume":"137","author":[{"given":"Daniel","family":"Basilio","sequence":"first","affiliation":[{"name":"Department of Physiology and Biophysics 1 and 2"},{"name":"Dominick P. Purpura Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461 1 and 2"},{"name":"Department of Physiology and Biophysics 1 and 2"},{"name":"Dominick P. Purpura Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461 1 and 2"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paul K.","family":"Kienker","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biophysics 1 and 2"},{"name":"Dominick P. Purpura Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461 1 and 2"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stephen W.","family":"Briggs","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biophysics 1 and 2"},{"name":"Dominick P. Purpura Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461 1 and 2"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alan","family":"Finkelstein","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biophysics 1 and 2"},{"name":"Dominick P. Purpura Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461 1 and 2"},{"name":"Department of Physiology and Biophysics 1 and 2"},{"name":"Dominick P. Purpura Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461 1 and 2"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"291","published-online":{"date-parts":[[2011,5,30]]},"reference":[{"key":"2023072921123724700_bib1","doi-asserted-by":"publisher","first-page":"307","DOI":"10.1085\/jgp.200810170","article-title":"Evidence for a proton-protein symport mechanism in the anthrax toxin channel","volume":"133","author":"Basilio","year":"2009","journal-title":"J. Gen. Physiol."},{"key":"2023072921123724700_bib2","doi-asserted-by":"publisher","first-page":"343","DOI":"10.1085\/jgp.201010578","article-title":"Trapping a translocating protein within the anthrax toxin channel: implications for the secondary structure of permeating proteins","volume":"137","author":"Basilio","year":"2011","journal-title":"J. Gen. Physiol."},{"key":"2023072921123724700_bib3","doi-asserted-by":"publisher","first-page":"3941","DOI":"10.1021\/bi972657b","article-title":"Identification of residues lining the anthrax protective antigen channel","volume":"37","author":"Benson","year":"1998","journal-title":"Biochemistry."},{"key":"2023072921123724700_bib4","doi-asserted-by":"publisher","first-page":"787","DOI":"10.1016\/S0006-3495(03)74898-X","article-title":"Translocation of rodlike polymers through membrane channels","volume":"84","author":"Berezhkovskii","year":"2003","journal-title":"Biophys. J."},{"key":"2023072921123724700_bib5","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1016\/0378-1119(89)90335-1","article-title":"Nucleotide sequence and analysis of the lethal factor gene (lef) from Bacillus anthracis","volume":"81","author":"Bragg","year":"1989","journal-title":"Gene."},{"key":"2023072921123724700_bib6","doi-asserted-by":"publisher","first-page":"7049","DOI":"10.1073\/pnas.062160399","article-title":"Mapping the lethal factor and edema factor binding sites on oligomeric anthrax protective antigen","volume":"99","author":"Cunningham","year":"2002","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072921123724700_bib7","doi-asserted-by":"publisher","first-page":"6706","DOI":"10.1021\/bi000310u","article-title":"A quantitative study of the interactions of Bacillus anthracis edema factor and lethal factor with activated protective antigen","volume":"39","author":"Elliott","year":"2000","journal-title":"Biochemistry."},{"key":"2023072921123724700_bib8","doi-asserted-by":"publisher","first-page":"209","DOI":"10.1098\/rstb.2008.0126","article-title":"Proton-coupled protein transport through the anthrax toxin channel","volume":"364","author":"Finkelstein","year":"2009","journal-title":"Philos. Trans. R. Soc. Lond. B Biol. Sci."},{"key":"2023072921123724700_bib9","doi-asserted-by":"publisher","first-page":"041910","DOI":"10.1103\/PhysRevE.68.041910","article-title":"Single stranded DNA translocation through a nanopore: a master equation approach","volume":"68","author":"Flomenbom","year":"2003","journal-title":"Phys. Rev. E Stat. Nonlin. Soft Matter Phys."},{"key":"2023072921123724700_bib10","doi-asserted-by":"publisher","first-page":"065103","DOI":"10.1063\/1.2826339","article-title":"A Monte Carlo algorithm to study polymer translocation through nanopores. I. Theory and numerical approach","volume":"128","author":"Gauthier","year":"2008","journal-title":"J. Chem. Phys."},{"key":"2023072921123724700_bib11","doi-asserted-by":"publisher","first-page":"1542","DOI":"10.1007\/s10773-006-9291-0","article-title":"Outflow probability for drift\u2013diffusion dynamics","volume":"46","author":"Hinkel","year":"2007","journal-title":"International Journal of Theoretical Physics."},{"key":"2023072921123724700_bib12","doi-asserted-by":"publisher","first-page":"754","DOI":"10.1038\/nsmb.1442","article-title":"GroEL as a molecular scaffold for structural analysis of the anthrax toxin pore","volume":"15","author":"Katayama","year":"2008","journal-title":"Nat. Struct. Mol. Biol."},{"key":"2023072921123724700_bib13","doi-asserted-by":"publisher","first-page":"614","DOI":"10.1016\/j.jmb.2009.07.037","article-title":"The protective antigen component of anthrax toxin forms functional octameric complexes","volume":"392","author":"Kintzer","year":"2009","journal-title":"J. Mol. Biol."},{"key":"2023072921123724700_bib14","doi-asserted-by":"publisher","first-page":"777","DOI":"10.1126\/science.1113380","article-title":"A phenylalanine clamp catalyzes protein translocation through the anthrax toxin pore","volume":"309","author":"Krantz","year":"2005","journal-title":"Science."},{"key":"2023072921123724700_bib15","doi-asserted-by":"publisher","first-page":"968","DOI":"10.1016\/j.jmb.2005.11.030","article-title":"Protein translocation through the anthrax toxin transmembrane pore is driven by a proton gradient","volume":"355","author":"Krantz","year":"2006","journal-title":"J. Mol. Biol."},{"key":"2023072921123724700_bib16","doi-asserted-by":"publisher","first-page":"1824","DOI":"10.1016\/S0006-3495(99)77027-X","article-title":"Driven polymer translocation through a narrow pore","volume":"77","author":"Lubensky","year":"1999","journal-title":"Biophys. J."},{"key":"2023072921123724700_bib17","doi-asserted-by":"publisher","first-page":"1630","DOI":"10.1074\/jbc.M511164200","article-title":"Structural determinants for the binding of anthrax lethal factor to oligomeric protective antigen","volume":"281","author":"Melnyk","year":"2006","journal-title":"J. Biol. Chem."},{"key":"2023072921123724700_bib18","doi-asserted-by":"publisher","first-page":"1079","DOI":"10.1021\/bi015860m","article-title":"Stoichiometry of anthrax toxin complexes","volume":"41","author":"Mogridge","year":"2002","journal-title":"Biochemistry."},{"key":"2023072921123724700_bib19","doi-asserted-by":"crossref","first-page":"534","DOI":"10.1021\/j100796a529","article-title":"Methods for the formation of single bimolecular lipid membranes in aqueous solutions","volume":"67","author":"Mueller","year":"1963","journal-title":"J. Phys. Chem."},{"key":"2023072921123724700_bib20","doi-asserted-by":"publisher","first-page":"1445","DOI":"10.1021\/bi0119518","article-title":"PA63 channel of anthrax toxin: an extended \u03b2-barrel","volume":"41","author":"Nassi","year":"2002","journal-title":"Biochemistry."},{"key":"2023072921123724700_bib21","doi-asserted-by":"publisher","first-page":"3060","DOI":"10.1021\/bi0524316","article-title":"Anthrax lethal factor (LF) mediated block of the anthrax protective antigen (PA) ion channel: effect of ionic strength and voltage","volume":"45","author":"Neumeyer","year":"2006","journal-title":"Biochemistry."},{"key":"2023072921123724700_bib22","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1080\/07391102.2004.10531226","article-title":"Three-dimensional model of the pore form of anthrax protective antigen. Structure and biological implications","volume":"22","author":"Nguyen","year":"2004","journal-title":"J. Biomol. Struct. Dyn."},{"key":"2023072921123724700_bib23","doi-asserted-by":"publisher","first-page":"258","DOI":"10.1016\/j.bbrc.2004.07.105","article-title":"Anthrax toxin complexes: heptameric protective antigen can bind lethal factor and edema factor simultaneously","volume":"322","author":"Pimental","year":"2004","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"2023072921123724700_bib24","doi-asserted-by":"publisher","first-page":"1859","DOI":"10.1126\/science.274.5294.1859","article-title":"Structure of staphylococcal alpha-hemolysin, a heptameric transmembrane pore","volume":"274","author":"Song","year":"1996","journal-title":"Science."},{"key":"2023072921123724700_bib25","doi-asserted-by":"publisher","first-page":"21555","DOI":"10.1073\/pnas.0905880106","article-title":"Lethal factor unfolding is the most force-dependent step of anthrax toxin translocation","volume":"106","author":"Thoren","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072921123724700_bib26","doi-asserted-by":"publisher","first-page":"031904","DOI":"10.1103\/PhysRevE.77.031904","article-title":"Orientation-dependent interactions of DNA with an \u03b1-hemolysin channel","volume":"77","author":"Wanunu","year":"2008","journal-title":"Phys. Rev. E Stat. Nonlin. Soft Matter Phys."},{"key":"2023072921123724700_bib27","doi-asserted-by":"publisher","first-page":"289","DOI":"10.1016\/S0006-3495(90)82376-6","article-title":"Optimizing planar lipid bilayer single-channel recordings for high resolution with rapid voltage steps","volume":"58","author":"Wonderlin","year":"1990","journal-title":"Biophys. J."},{"key":"2023072921123724700_bib28","doi-asserted-by":"publisher","first-page":"045101","DOI":"10.1063\/1.3464333","article-title":"Polymer translocation through alpha-hemolysin pore with tunable polymer-pore electrostatic interaction","volume":"133","author":"Wong","year":"2010","journal-title":"J. Chem. Phys."},{"key":"2023072921123724700_bib29","doi-asserted-by":"publisher","first-page":"243","DOI":"10.1146\/annurev.biochem.75.103004.142728","article-title":"Anthrax toxin: receptor binding, internalization, pore formation, and translocation","volume":"76","author":"Young","year":"2007","journal-title":"Annu. Rev. Biochem."},{"key":"2023072921123724700_bib30","doi-asserted-by":"publisher","first-page":"16756","DOI":"10.1073\/pnas.0405754101","article-title":"Evidence that translocation of anthrax toxin\u2019s lethal factor is initiated by entry of its N terminus into the protective antigen channel","volume":"101","author":"Zhang","year":"2004","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072921123724700_bib31","doi-asserted-by":"publisher","first-page":"3842","DOI":"10.1529\/biophysj.104.050864","article-title":"Protein translocation through anthrax toxin channels formed in planar lipid bilayers","volume":"87","author":"Zhang","year":"2004","journal-title":"Biophys. J."}],"container-title":["Journal of General Physiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/rupress.org\/jgp\/article-pdf\/137\/6\/521\/1914517\/jgp_201110627.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/rupress.org\/jgp\/article-pdf\/137\/6\/521\/1914517\/jgp_201110627.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,7,29]],"date-time":"2023-07-29T21:13:09Z","timestamp":1690665189000},"score":1,"resource":{"primary":{"URL":"https:\/\/rupress.org\/jgp\/article\/137\/6\/521\/42983\/A-kinetic-analysis-of-protein-transport-through"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2011,5,30]]},"references-count":31,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2011,6,1]]}},"URL":"https:\/\/doi.org\/10.1085\/jgp.201110627","relation":{},"ISSN":["1540-7748","0022-1295"],"issn-type":[{"value":"1540-7748","type":"electronic"},{"value":"0022-1295","type":"print"}],"subject":[],"published":{"date-parts":[[2011,5,30]]}}}