{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2024,8,10]],"date-time":"2024-08-10T05:57:00Z","timestamp":1723269420655},"reference-count":26,"publisher":"Rockefeller University Press","issue":"4","content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2011,4,1]]},"abstract":"<jats:p>Anthrax toxin consists of three proteins: lethal factor (LF), edema factor (EF), and protective antigen (PA). This last forms a heptameric channel, (PA63)7, in the host cell\u2019s endosomal membrane, allowing the former two (which are enzymes) to be translocated into the cytosol. (PA63)7 incorporated into planar bilayer membranes forms a channel that translocates LF and EF, with the N terminus leading the way. The channel is mushroom-shaped with a cap containing the binding sites for EF and LF, and an \u223c100 \u00c5\u2013long, 15 \u00c5\u2013wide stem. For proteins to pass through the stem they clearly must unfold, but is secondary structure preserved? To answer this question, we developed a method of trapping the polypeptide chain of a translocating protein within the channel and determined the minimum number of residues that could traverse it. We attached a biotin to the N terminus of LFN (the 263-residue N-terminal portion of LF) and a molecular stopper elsewhere. If the distance from the N terminus to the stopper was long enough to traverse the channel, streptavidin added to the trans side bound the N-terminal biotin, trapping the protein within the channel; if this distance was not long enough, streptavidin did not bind the N-terminal biotin and the protein was not trapped. The trapping rate was dependent on the driving force (voltage), the length of time it was applied, and the number of residues between the N terminus and the stopper. By varying the position of the stopper, we determined the minimum number of residues required to span the channel. We conclude that LFN adopts an extended-chain configuration as it translocates; i.e., the channel unfolds the secondary structure of the protein. We also show that the channel not only can translocate LFN in the normal direction but also can, at least partially, translocate LFN in the opposite direction.<\/jats:p>","DOI":"10.1085\/jgp.201010578","type":"journal-article","created":{"date-parts":[[2011,3,15]],"date-time":"2011-03-15T18:54:13Z","timestamp":1300215253000},"page":"343-356","update-policy":"http:\/\/dx.doi.org\/10.1085\/jgp.crossmarkpolicy","source":"Crossref","is-referenced-by-count":29,"title":["Trapping a translocating protein within the anthrax toxin channel: implications for the secondary structure of permeating proteins"],"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":"Laura D.","family":"Jennings-Antipov","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115 3"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Karen S.","family":"Jakes","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":[[2011,3,14]]},"reference":[{"key":"2023072521322276200_bib1","doi-asserted-by":"publisher","first-page":"351","DOI":"10.1085\/jgp.200809984","article-title":"Preventing voltage-dependent gating of anthrax toxin channels using engineered disulfides","volume":"132","author":"Anderson","year":"2008","journal-title":"J. Gen. 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