Analyzing topography of membrane-inserted diphtheria toxin T domain using BODIPY-streptavidin: at low pH, helices 8 and 9 form a transmembrane hairpin but helices 5-7 form stable nonclassical inserted segments on the cis side of the bilayer

Biochemistry
Michael P RosconiE London

Abstract

Low pH-induced membrane insertion by diphtheria toxin T domain is crucial for A chain translocation into the cytoplasm. To define the membrane topography of the T domain, the exposure of biotinylated Cys residues to the cis and trans bilayer surfaces was examined using model membrane vesicles containing a deeply inserted T domain. To do this, the reactivity of biotin with external and vesicle-entrapped BODIPY-labeled streptavidin was measured. The T domain was found to insert with roughly 70-80% of the molecules in the physiologically relevant orientation. In this orientation, residue 349, located in the loop between hydrophobic helices 8 and 9, was exposed to the trans side of the bilayer, while other solution-exposed residues along the hydrophobic helices 5-9 region of the T domain located near the cis surface. A protocol developed to detect the movement of residues back and forth across the membranes demonstrated that T domain sequences did not rapidly equilibrate between the cis and the trans sides of the bilayer. Binding streptavidin to biotinylated residues prior to membrane insertion only inhibited T domain pore formation for residues in the loop between helices 8 and 9. Pore formation experiments used an approach avoidi...Continue Reading

References

Dec 1, 1980·The Journal of Cell Biology·K Sandvig, S Olsnes
Jan 1, 1994·Molecular Microbiology·V CabiauxJ M Ruysschaert

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Citations

Mar 22, 2007·European Biophysics Journal : EBJ·Valery G Veresov, Alexander I Davidovskii
Nov 22, 2005·Analytical Biochemistry·Yevgen O Posokhov, Alexey S Ladokhin
Apr 9, 2005·Gene Therapy·K Sandvig, B van Deurs
Apr 30, 2013·Biochemistry·Mauricio Vargas-UribeAlexey S Ladokhin
Dec 10, 2015·The Journal of Membrane Biology·Paul K KienkerAlan Finkelstein
Oct 18, 2006·Biology of the Cell·Blandine Geny, Michel R Popoff
Mar 19, 2014·The FEBS Journal·Oneda LekaGiuseppe Zanotti
Mar 2, 2005·Biochemical and Biophysical Research Communications·Bjørn SpilsbergKirsten Sandvig
Oct 5, 2014·The Journal of Membrane Biology·Mauricio Vargas-UribeAlexey S Ladokhin
Apr 9, 2015·The Journal of Membrane Biology·Qingqing LinErwin London
Nov 2, 2005·Toxicon : Official Journal of the International Society on Toxinology·Bjørn SpilsbergSébastien Wälchli
Jan 15, 2015·The Journal of General Physiology·Paul K KienkerAlan Finkelstein
May 9, 2013·Biochemistry·Smita RaghavaDaniel N Hebert
May 27, 2017·Critical Reviews in Biochemistry and Molecular Biology·Kathleen E OrrellRoman A Melnyk
Jan 9, 2014·The Journal of Biological Chemistry·Qingqing Lin, Erwin London
Jan 21, 2016·The Journal of Biological Chemistry·Fabian B RomanoAlejandro P Heuck
Apr 21, 2019·Biochimica Et Biophysica Acta. Proteins and Proteomics·Victor Vasquez-MontesAlexey S Ladokhin
Feb 9, 2005·Biochemistry·Masatoshi Hayashibara, Erwin London
May 11, 2006·Chemical Reviews·Kevin R Mackenzie

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