Home LiteratureArticle Details
PMID: 7068762 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Isolation of intracellular membranes by means of sodium carbonate treatment: application to endoplasmic reticulum.

The Journal of cell biology ·Vol. 93 ·No. 1 ·1982-04-00 ·Pages 97-102

Fujiki Y, Hubbard AL, Fowler S, Lazarow PB

Abstract

A rapid and simple method for the isolation of membranes from subcellular organelles is described. The procedure consists of diluting the organelles in ice-cold 100 mM Na2CO3 followed by centrifugation to pellet the membranes. Closed vesicles are converted to open membrane sheets, and content proteins and peripheral membrane proteins are released in soluble form. Here we document the method by applying it to various subfractions of a rat liver microsomal fraction, prepared by continuous density gradient centrifugation according to Beaufay et al. (1974, J. Cell Biol. 61:213-231). The results confirm and extend those of previous investigators on the distribution of enzymes and proteins among the membranes of the smooth and rough endoplasmic reticulum. In the accompanying paper (1982, J. Cell Biol. 93:103-110) the procedure is applied to peroxisomes and mitochondria.

MeSH Terms
Animals Carbonates Cell Fractionation/methods Centrifugation, Density Gradient/methods Female Intracellular Membranes/ultrastructure Liver/ultrastructure Microscopy, Electron Microsomes, Liver/ultrastructure Rats Rats, Inbred Strains Sarcoplasmic Reticulum/ultrastructure
Chemicals
Carbonates sodium carbonate
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Fujiki Y
Hubbard A L
Fowler S
Lazarow P B
References (27)
27 references, click to expand
  1. Tissue fractionation studies. 18. Resolution of mitochondrial fractions from rat liver into three distinct populations of cytoplasmic particles by means of density equilibration in various gradients.
    Biochem J. 1964 Jul;92(1):184-205 PMID: 4378797
  2. Cell junctions in amphibian skin.
    J Cell Biol. 1965 Jul;26(1):263-91 PMID: 5859021
  3. The large-scale separation of peroxisomes, mitochondria, and lysosomes from the livers of rats injected with triton WR-1339. Improved isolation procedures, automated analysis, biochemical and morphological properties of fractions.
    J Cell Biol. 1968 May;37(2):482-513 PMID: 4297786
  4. Distribution of radioactivity between the acid-soluble pool and the pools of RNA in the nuclear, nonsedimethable and ribosome fractions of rat liver after a single injection of lebaled orotic acid.
    Biochim Biophys Acta. 1968 Aug 23;166(1):48-57 PMID: 5698065
  5. Solubility of membrane proteins in aqueous media.
    J Biol Chem. 1971 Dec 10;246(23):7337-42 PMID: 5002002
  6. The fluid mosaic model of the structure of cell membranes.
    Science. 1972 Feb 18;175(4023):720-31 PMID: 4333397
  7. Ribosome-membrane interaction. Nondestructive disassembly of rat liver rough microsomes into ribosomal and membranous components.
    J Cell Biol. 1973 Jan;56(1):206-29 PMID: 4682341
  8. Lipid-protein interactions in the reconstitution of the microsomal reduced nicotinamide adenine dinucleotide-cytochrome b 5 reductase system.
    J Biol Chem. 1973 Feb 10;248(3):800-6 PMID: 4346351
  9. Selective release of content from microsomal vesicles without membrane disassembly. I. Permeability changes induced by low detergent concentrations.
    J Cell Biol. 1973 Aug;58(2):436-62 PMID: 4729506
  10. Synthesis of rat liver albumin in a rabbit reticulocyte cell-free protein-synthesizing system.
    J Biol Chem. 1973 Nov 25;248(22):7661-8 PMID: 4750421
  11. Analytical study of microsomes and isolated subcellular membranes from rat liver. I. Biochemical methods.
    J Cell Biol. 1974 Apr;61(1):188-200 PMID: 4150488
  12. Analytical study of microsomes and isolated subcellular membranes from rat liver. 3. Subfractionation of the microsomal fraction by isopycnic and differential centrifugation in density gradients.
    J Cell Biol. 1974 Apr;61(1):213-31 PMID: 4150490
  13. Selective release of content from microsomal vesicles without membrane disassembly. II. Electrophoretic and immunological characterization of microsomal subfractions.
    J Cell Biol. 1974 Jun;61(3):789-807 PMID: 4209578
  14. The organization of proteins in the human red blood cell membrane. A review.
    J Cell Biol. 1974 Jul;62(1):1-19 PMID: 4600883
  15. Analytical study of microsomes and isolated subcellular membranes from rat liver. IV. Biochemical, physical, and morphological modifications of microsomal components induced by digitonin, EDTA, and pyrophosphate.
    J Cell Biol. 1974 Sep;62(3):717-45 PMID: 4368410
  16. Selective solubilization of proteins from red blood cell membranes by protein perturbants.
    J Supramol Struct. 1973;1(3):220-32 PMID: 4804837
  17. The synthesis and turnover of rat liver peroxisomes. V. Intracellular pathway of catalase synthesis.
    J Cell Biol. 1973 Nov;59(2 Pt 1):507-24 PMID: 4805011
  18. Proteins of rough microsomal membranes related to ribosome binding. I. Identification of ribophorins I and II, membrane proteins characteristics of rough microsomes.
    J Cell Biol. 1978 May;77(2):464-87 PMID: 649658
  19. Cell fractionation studies on the guinea pig pancreas. Redistribution of exocrine proteins during tissue homogenization.
    J Cell Biol. 1978 Jul;78(1):110-30 PMID: 670290
  20. Synthesis of catalase in two cell-free protein-synthesizing systems and in rat liver.
    Proc Natl Acad Sci U S A. 1978 Sep;75(9):4344-8 PMID: 279920
  21. Characterization of the ribosomal binding site in rat liver rough microsomes: ribophorins I and II, two integral membrane proteins related to ribosome binding.
    J Supramol Struct. 1978;8(3):279-302 PMID: 723266
  22. Synthesis of rat liver microsomal cytochrome b5 by free ribosomes.
    J Cell Biol. 1980 Mar;84(3):705-16 PMID: 7358795
  23. Polypeptide and phospholipid composition of the membrane of rat liver peroxisomes: comparison with endoplasmic reticulum and mitochondrial membranes.
    J Cell Biol. 1982 Apr;93(1):103-10 PMID: 7068748
  24. Protein measurement with the Folin phenol reagent.
    J Biol Chem. 1951 Nov;193(1):265-75 PMID: 14907713
  25. A rapid method of total lipid extraction and purification.
    Can J Biochem Physiol. 1959 Aug;37(8):911-7 PMID: 13671378
  26. Tissue fractionation studies. 6. Intracellular distribution patterns of enzymes in rat-liver tissue.
    Biochem J. 1955 Aug;60(4):604-17 PMID: 13249955
  27. The role of polyamines in the neutralization of bacteriophage deoxyribonucleic acid.
    J Biol Chem. 1960 Mar;235:769-75 PMID: 13793161
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.93.1.97
Published
1982-04-00
Pages
97-102
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2112113
Subset
IM
Grants
NIADDK NIH HHS · AM 19394 · United States
NIGMS NIH HHS · GM 29185 · United States
External Links
PubMed source DOI: 10.1083/jcb.93.1.97 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2