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  Transients of perforin pore formation observed by fluorescence microscopic single channel recording

Peters, R., Sauer, H., Tschopp, J., & Fritzsch, G. (1990). Transients of perforin pore formation observed by fluorescence microscopic single channel recording. EMBO Journal, 9(8), 2447-2451. doi:10.1002/j.1460-2075.1990.tb07421.x.

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 Creators:
Peters, Reiner1, 2, Author              
Sauer, Heinrich1, Author              
Tschopp, Jürgen3, Author
Fritzsch, Günter4, Author              
Affiliations:
1Department of Cell Physiology, Max Planck Institute of Biophysics, Max Planck Society, ou_3264817              
2Institute for Medical Physics, Westfälische Wilhelms-Universität, D-4400 Münster, Germany, ou_persistent22              
3Institute of Biochemistry, University of Lausanne, CH-1066 Epalinges, Switzerland, ou_persistent22              
4Department of Molecular Membrane Biology, Max Planck Institute of Biophysics, Max Planck Society, ou_2068290              

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Free keywords: cytolysin; erythrocyte membrane; perforin; photo-bleaching; single channel recording
 Abstract: A new type of single channel recording is described. Large pores were generated in the membranes of resealed human erythrocyte ghosts by incubation with perforin (cytolysin). The flux of the polar fluorescent probe Lucifer Yellow was measured in single ghosts by the fluorescence microphotolysis (photobleaching) technique. The distribution of flux rates for ghosts treated with a limiting perforin concentration showed equidistantly spaced peaks suggesting that subpopulations of ghosts with 0, 1 and 2 pores were resolved. Furthermore, distributions obtained for very different perforin concentrations could be well simulated by using one common value for the flux rate of the single pore (k = 4.65 x 10-3s) and assuming a Poisson distribution of pores among ghosts. The flux rate of the single pore corresponds to a pore radius of approximately 50 A, a value which is much smaller than that obtained previously by electron microscopic studies but which agrees well with recent electrical single channel recordings. Mature perforin pores were observed to be very stable. No closing events were detected at a time resolution of 0.2 s for a wide range of temperatures and Ca2+ concentrations. However, the formation of new pores was an unexpectedly slow process. Fluorescence microscopic single channel recording as introduced by this study is applicable to a variety of cellular systems and fluorescent probes and thus may complement the information obtainable by electrical single channel recording of anorganic ion fluxes

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Language(s): eng - English
 Dates: 1990-04-301990-03-231990-04-301990-08-01
 Publication Status: Published in print
 Pages: 5
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1002/j.1460-2075.1990.tb07421.x
PMID: 1695147
 Degree: -

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Title: EMBO Journal
  Other : EMBO J.
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: Nature Publishing Group
Pages: - Volume / Issue: 9 (8) Sequence Number: - Start / End Page: 2447 - 2451 Identifier: ISSN: 0261-4189
CoNE: https://pure.mpg.de/cone/journals/resource/954925497061