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. 1973 Aug 1;58(2):436–462. doi: 10.1083/jcb.58.2.436

SELECTIVE RELEASE OF CONTENT FROM MICROSOMAL VESICLES WITHOUT MEMBRANE DISASSEMBLY

I. Permeability Changes Induced by Low Detergent Concentrations

Gert Kreibich 1, Pascale Debey 1, David D Sabatini 1
PMCID: PMC2109052  PMID: 4729506

Abstract

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.

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Selected References

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  1. AMES B. N., DUBIN D. T. The role of polyamines in the neutralization of bacteriophage deoxyribonucleic acid. J Biol Chem. 1960 Mar;235:769–775. [PubMed] [Google Scholar]
  2. Adelman M. R., Blobel G., Sabatini D. D. An improved cell fractionation procedure for the preparation of rat liver membrane-bound ribosomes. J Cell Biol. 1973 Jan;56(1):191–205. doi: 10.1083/jcb.56.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Adelman M. R., Sabatini D. D., Blobel G. Ribosome-membrane interaction. Nondestructive disassembly of rat liver rough microsomes into ribosomal and membranous components. J Cell Biol. 1973 Jan;56(1):206–229. doi: 10.1083/jcb.56.1.206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Agostini B., Hasselbach W. Influence of lysolecithin on fragmented sarcoplasmic reticulum. Naturwissenschaften. 1971 Mar;58(3):148–148. doi: 10.1007/BF00593107. [DOI] [PubMed] [Google Scholar]
  5. Benzonana G. Study of bile salts micelles: properties of mixed oleate-deoxycholate solutions at pH 9.0. Biochim Biophys Acta. 1969 Jun 10;176(4):836–848. doi: 10.1016/0005-2760(69)90265-3. [DOI] [PubMed] [Google Scholar]
  6. Borgese D., Blobel G., Sabatini D. D. In vitro exchange of ribosomal subunits between free and membrane-bound ribosomes. J Mol Biol. 1973 Mar 15;74(4):415–438. doi: 10.1016/0022-2836(73)90037-5. [DOI] [PubMed] [Google Scholar]
  7. CAMPBELL P. N., GREENGARD O., KERNOT B. A. Studies on the synthesis o serum albumin by the isolated microsome fraction from rat liver. Biochem J. 1960 Jan;74:107–117. doi: 10.1042/bj0740107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Colbeau A., Nachbaur J., Vignais P. M. Enzymic characterization and lipid composition of rat liver subcellular membranes. Biochim Biophys Acta. 1971 Dec 3;249(2):462–492. doi: 10.1016/0005-2736(71)90123-4. [DOI] [PubMed] [Google Scholar]
  9. Dallman P. R., Dallner G., Bergstrand A., Ernster L. Heterogeneous distribution of enzymes in submicrosomal membrane fragments. J Cell Biol. 1969 May;41(2):357–377. doi: 10.1083/jcb.41.2.357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. De Lorenzo F., Goldberger R. F., Steers E., Jr, Givol D., Anfinsen B. Purification and properties of an enzyme from beef liver which catalyzes sulfhydryl-disulfide interchange in proteins. J Biol Chem. 1966 Apr 10;241(7):1562–1567. [PubMed] [Google Scholar]
  11. Engelman D. M., Terry T. M., Morowitz H. J. Characterization of the plasma membrane of Mycoplasma laidlawii. I. Sodium dodecyl sulfate solubilization. Biochim Biophys Acta. 1967 Jul 3;135(3):381–390. doi: 10.1016/0005-2736(67)90028-4. [DOI] [PubMed] [Google Scholar]
  12. Estrugo S. F., Larraga V., Corrales M. A., Duch C., Muñoz E. Molecular organization in bacterial cell membranes. I. Sodium dodecyl sulphate solubilization and fractionation of the components of a depleted membrane from Micrococcus lysodeikticus. Biochim Biophys Acta. 1972 Mar 17;255(3):960–973. doi: 10.1016/0005-2736(72)90406-3. [DOI] [PubMed] [Google Scholar]
  13. FOLCH J., LEES M., SLOANE STANLEY G. H. A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem. 1957 May;226(1):497–509. [PubMed] [Google Scholar]
  14. GARFINKEL D. Preparation and properties of a microsomal DPNH-cytochrome c reductase. Arch Biochem Biophys. 1957 Sep;71(1):100–110. doi: 10.1016/0003-9861(57)90011-5. [DOI] [PubMed] [Google Scholar]
  15. Glaumann H., Ericsson J. L. Evidence for the participation of the Golgi apparatus in the intracellular transport of nascent albumin in the liver cell. J Cell Biol. 1970 Dec;47(3):555–567. doi: 10.1083/jcb.47.3.555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Glaumann H. Studies on the synthesis and transport of albumin in microsomal subfractions from rat liver. Biochim Biophys Acta. 1970 Nov 12;224(1):206–218. doi: 10.1016/0005-2787(70)90634-9. [DOI] [PubMed] [Google Scholar]
  17. HODES M. E., PALMER C. G., WARREN A. The effect of surface active agents on the permeability to dye of the plasma membrane of Ehrlich ascites cells. Exp Cell Res. 1960 Oct;21:164–169. doi: 10.1016/0014-4827(60)90357-8. [DOI] [PubMed] [Google Scholar]
  18. Hall J. D., Crane F. L. Fractionation of mitochondrial membranes with sodium deoxycholate. Biochim Biophys Acta. 1972 Feb 11;255(2):602–619. doi: 10.1016/0005-2736(72)90164-2. [DOI] [PubMed] [Google Scholar]
  19. Ito A., Sato R. Proteolytic microdissection of smooth-surfaced vesicles of liver microsomes. J Cell Biol. 1969 Jan;40(1):179–189. doi: 10.1083/jcb.40.1.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kuriyama Y. Studies on microsomal nucleoside diphosphatase of rat hepatocytes. Its purification, intramembranous localization, and turnover. J Biol Chem. 1972 May 25;247(10):2979–2988. [PubMed] [Google Scholar]
  21. LUST J., DROCHMANS P. Effect of trypsin on liver microsomes. J Cell Biol. 1963 Jan;16:81–92. doi: 10.1083/jcb.16.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lee S. Y., Krsmanovic V., Brawerman G. Attachment of ribosomes to membranes during polysome formation in mouse sarcoma 180 cells. J Cell Biol. 1971 Jun;49(3):683–691. doi: 10.1083/jcb.49.3.683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lucy J. A. The fusion of biological membranes. Nature. 1970 Aug 22;227(5260):815–817. doi: 10.1038/227815a0. [DOI] [PubMed] [Google Scholar]
  24. Malenkov A. G., Bogatyreva S. A., Bozhkova V. P., Modjanova E. A., Vasiliev J. M. Reversible alterations of the surface of ascites tumour cells induced by a surface-active substance, Tween 60. Exp Cell Res. 1967 Nov;48(2):307–318. doi: 10.1016/0014-4827(67)90356-4. [DOI] [PubMed] [Google Scholar]
  25. Ne'eman Z., Kahane I., Razin S. Characterization of the mycoplasma membrane proteins. II. Solubilization and enzymic activities of Acholeplasma laidlawii membrane proteins. Biochim Biophys Acta. 1971 Oct 12;249(1):169–176. doi: 10.1016/0005-2736(71)90093-9. [DOI] [PubMed] [Google Scholar]
  26. Omura T., Siekevitz P., Palade G. E. Turnover of constituents of the endoplasmic reticulum membranes of rat hepatocytes. J Biol Chem. 1967 May 25;242(10):2389–2396. [PubMed] [Google Scholar]
  27. PALADE G. E., SIEKEVITZ P. Liver microsomes; an integrated morphological and biochemical study. J Biophys Biochem Cytol. 1956 Mar 25;2(2):171–200. doi: 10.1083/jcb.2.2.171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Peters T., Jr, Fleischer B., Fleischer S. The biosynthesis of rat serum albumin. IV. Apparent passage of albumin through the Golgi apparatus during secretion. J Biol Chem. 1971 Jan 10;246(1):240–244. [PubMed] [Google Scholar]
  29. Philippot J. Study of human red blood cell membrane using sodium deoxycholate. I. Mechanism of the solubilization. Biochim Biophys Acta. 1971 Feb 2;225(2):201–213. doi: 10.1016/0005-2736(71)90213-6. [DOI] [PubMed] [Google Scholar]
  30. Poole A. R., Howell J. I., Lucy J. A. Lysolecithin and cell fusion. Nature. 1970 Aug 22;227(5260):810–814. doi: 10.1038/227810a0. [DOI] [PubMed] [Google Scholar]
  31. ROTH J. S. Studies on the function of intracellular ribonucleases. II. The interaction of ribonucleprotein and enzymes. J Biophys Biochem Cytol. 1960 Jun;7:443–453. doi: 10.1083/jcb.7.3.443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Redman C. M. Biosynthesis of serum proteins and ferritin by free and attached ribosomes of rat liver. J Biol Chem. 1969 Aug 25;244(16):4308–4315. [PubMed] [Google Scholar]
  33. Redman C. M., Cherian M. G. The secretory pathways of rat serum glycoproteins and albumin. Localization of newly formed proteins within the endoplasmic reticulum. J Cell Biol. 1972 Feb;52(2):231–245. doi: 10.1083/jcb.52.2.231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Redman C. M., Sabatini D. D. Vectorial discharge of peptides released by puromycin from attached ribosomes. Proc Natl Acad Sci U S A. 1966 Aug;56(2):608–615. doi: 10.1073/pnas.56.2.608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Redman C. M., Siekevitz P., Palade G. E. Synthesis and transfer of amylase in pigeon pancreatic micromosomes. J Biol Chem. 1966 Mar 10;241(5):1150–1158. [PubMed] [Google Scholar]
  36. Redman C. M. Studies on the transfer of incomplete polypeptide chains across rat liver microsomal membranes in vitro. J Biol Chem. 1967 Feb 25;242(4):761–768. [PubMed] [Google Scholar]
  37. Rostgaard J., Moller O. J. Electron microscopy of a microsomal fraction ritch in (Na++K+)-ATPase and isolated from kidney cortex. Structural changes accompanying freeze- and DOC-activation and identification of main components. Exp Cell Res. 1971 Oct;68(2):356–371. doi: 10.1016/0014-4827(71)90161-3. [DOI] [PubMed] [Google Scholar]
  38. Sabatini D. D., Blobel G. Controlled proteolysis of nascent polypeptides in rat liver cell fractions. II. Location of the polypeptides in rough microsomes. J Cell Biol. 1970 Apr;45(1):146–157. doi: 10.1083/jcb.45.1.146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Schachter H., Jabbal I., Hudgin R. L., Pinteric L., McGuire E. J., Roseman S. Intracellular localization of liver sugar nucleotide glycoprotein glycosyltransferases in a Golgi-rich fraction. J Biol Chem. 1970 Mar 10;245(5):1090–1100. [PubMed] [Google Scholar]
  40. Seeman P., Cheng D., Iles G. H. Structure of membrane holes in osmotic and saponin hemolysis. J Cell Biol. 1973 Feb;56(2):519–527. doi: 10.1083/jcb.56.2.519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Seeman P. Transient holes in the erythrocyte membrane during hypotonic hemolysis and stable holes in the membrane after lysis by saponin and lysolecithin. J Cell Biol. 1967 Jan;32(1):55–70. doi: 10.1083/jcb.32.1.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sha'afi R. I., Dakkuri A., To'mey G. Solute and solvent flow across mammalian red cell membrane. How to test for Onsager reciprocal relation. Biochim Biophys Acta. 1971 Oct 12;249(1):260–265. doi: 10.1016/0005-2736(71)90103-9. [DOI] [PubMed] [Google Scholar]
  43. Siekevitz P. Biological membranes: the dynamics of their organization. Annu Rev Physiol. 1972;34:117–140. doi: 10.1146/annurev.ph.34.030172.001001. [DOI] [PubMed] [Google Scholar]
  44. Singer S. J., Nicolson G. L. The fluid mosaic model of the structure of cell membranes. Science. 1972 Feb 18;175(4023):720–731. doi: 10.1126/science.175.4023.720. [DOI] [PubMed] [Google Scholar]
  45. Steck T. L., Fairbanks G., Wallach D. F. Disposition of the major proteins in the isolated erythrocyte membrane. Proteolytic dissection. Biochemistry. 1971 Jun 22;10(13):2617–2624. doi: 10.1021/bi00789a031. [DOI] [PubMed] [Google Scholar]
  46. TASHIRO Y., SIEKEVITZ P. ULTRACENTRIFUGAL STUDIES ON THE DISSOCIATION OF HEPATIC RIBOSOMES. J Mol Biol. 1965 Feb;11:149–165. doi: 10.1016/s0022-2836(65)80047-x. [DOI] [PubMed] [Google Scholar]
  47. Tsukagoshi N., Fox C. F. Hybridization of membranes by sonic irradiation. Biochemistry. 1971 Aug 17;10(17):3309–3313. doi: 10.1021/bi00793a023. [DOI] [PubMed] [Google Scholar]
  48. WALLACH D. F., KAMAT V. B. PLASMA AND CYTOPLASMIC MEMBRANE FRAGMENTS FROM EHRLICH ASCITES CARCINOMA. Proc Natl Acad Sci U S A. 1964 Sep;52:721–728. doi: 10.1073/pnas.52.3.721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Weihing R. R., Manganiello V. C., Chiu R., Phillips A. H. Purification of hepatic microsomal membranes. Biochemistry. 1972 Aug 1;11(16):3128–3135. doi: 10.1021/bi00766a028. [DOI] [PubMed] [Google Scholar]

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