Rat liver rough microsomes treated with a series of desoxycholate (DOC) concentrations from 0.003 to 0.4% were analyzed by isopycnic sucrose density gradient centrifugation in media containing high or low salt concentrations. Tritium-labeled precursors administered in vivo were used as markers for ribosomes (orotic acid, 40 h), phospholipids (choline, 4 h), membrane proteins (leucine, 3 days), and completed secretory proteins of the vesicular cavity (leucine, 30 min). Within a narrow range of DOC concentrations (0.025–0.05%), the vesicular polypeptides were selectively released from the microsomes, while ribosomes, nascent polypeptides, and microsomal enzymes of the electron transport systems were unaffected. The detergent concentration which led to leakage of content was a function of the ionic strength and of the microsome concentration. At the lowest effective DOC concentration the microsomal membranes became reversibly permeable to macromoles as shown by changes in the density of the vesicles in Dextran gradients and by the extent of proteolysis by added proteases. Incubation of rough microsomes with proteases in the presence of 0.025% DOC also led to digestion of proteins from both faces of the microsomal membranes and to a lighter isopycnic density of the membrane vesicles.
Article|
August 01 1973
SELECTIVE RELEASE OF CONTENT FROM MICROSOMAL VESICLES WITHOUT MEMBRANE DISASSEMBLY : I. Permeability Changes Induced by Low Detergent Concentrations
Gert Kreibich,
Gert Kreibich
From The Rockefeller University, New York 10021 and New York University School of Medicine, New York 10016.
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Pascale Debey,
Pascale Debey
From The Rockefeller University, New York 10021 and New York University School of Medicine, New York 10016.
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David D. Sabatini
David D. Sabatini
From The Rockefeller University, New York 10021 and New York University School of Medicine, New York 10016.
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Gert Kreibich
From The Rockefeller University, New York 10021 and New York University School of Medicine, New York 10016.
Pascale Debey
From The Rockefeller University, New York 10021 and New York University School of Medicine, New York 10016.
David D. Sabatini
From The Rockefeller University, New York 10021 and New York University School of Medicine, New York 10016.
P. Debey's present address is Institut de Biologie Physico Chemique, Foundation de Rothschild, Paris 5e, France.
Received:
March 12 1973
Revision Received:
April 06 1973
Online ISSN: 1540-8140
Print ISSN: 0021-9525
Copyright © 1973 by The Rockefeller University Press
1973
J Cell Biol (1973) 58 (2): 436–462.
Article history
Received:
March 12 1973
Revision Received:
April 06 1973
Citation
Gert Kreibich, Pascale Debey, David D. Sabatini; SELECTIVE RELEASE OF CONTENT FROM MICROSOMAL VESICLES WITHOUT MEMBRANE DISASSEMBLY : I. Permeability Changes Induced by Low Detergent Concentrations . J Cell Biol 1 August 1973; 58 (2): 436–462. doi: https://doi.org/10.1083/jcb.58.2.436
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