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Distribution analysis of membrane penetration of proteins by depth-dependent fluorescence quenching

  • Quenching of Fluorescence
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Abstract

A new approach is presented to evaluate the depth-dependent quenching of the fluorescence of membrane-bound probes and integral proteins. By utilizing at least three quenchers of known and distinctly different depths, the following parameters can be recovered: most probable depth of the probe; dispersion of the depth distribution, which will depend on the size of probe and fluctuations in its position; and quenching efficiency, which is related to the exposure of a particular fluorophore to the lipid phase. The exposure of tryptophan residues in integral proteins can be quantitatively determined with respect to the model compound (tryptophan octyl ester). The proposed method was applied to the investigation of membrane complexes of the bee venom melittin and cytochrome b5.

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Authors and Affiliations

  1. Biology Department and McCollum-Pratt Institute, The Johns Hopkins University, 21218, Baltimore, Maryland

    Alexey S. Ladokhin

  2. Department of Biochemistry, University of Virginia School of Medicine, 22908, Charlottesville, Virginia

    Alexey S. Ladokhin & P. W. Holloway

  3. A. V. Palladin Institute of Biochemistry, Ukrainian Academy of Sciences, Kiev, Ukraine

    Alexey S. Ladokhin & E. G. Kostrzhevska

Authors
  1. Alexey S. Ladokhin
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  2. P. W. Holloway
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  3. E. G. Kostrzhevska
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Ladokhin, A.S., Holloway, P.W. & Kostrzhevska, E.G. Distribution analysis of membrane penetration of proteins by depth-dependent fluorescence quenching. J Fluoresc 3, 195–197 (1993). https://doi.org/10.1007/BF00862742

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  • DOI: https://doi.org/10.1007/BF00862742

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