Rapid Isolation of Plasma Membranes in High Yield from Cultured Fibroblasts

Overview

Journal Biochem J

Specialty Biochemistry

Date 1977 Nov 15

PMID 202261

Citations 59

Authors

D Thom

A J Powell

C W Lloyd

D A Rees

Affiliations

Soon will be listed here.

Abstract

1. A method was developed which allows the rapid preparation of pure plasma membranes in high yield from cultured fibroblasts. 2. Cells are lysed in hypo-osmotic borate/EDTA and, after differential centrifugation, the membranes collected by centrifugation on a sucrose barrier. 3. Electron microscopy of the isolated material shows large membrane vesicles essentially free from contaminating organelles. 4. There is no detectable activity of the endoplasmic-reticulum enzyme marker, NADH2--lipoamide oxidoreductase (EC 1.6.4.3), and that of succinate dehydrogenase (EC 1.3.99.1), a marker for mitochondria, is substantially decreased. Chemical compositions are in good agreement with previous observations. 5. This study confirms the usefulness of applied isotopic markers for isolating plasma membranes.

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References

Juliano R, Gagalang E . The isolation and characterization of the plasma membrane of cultured chinese hamster ovary cells. Can J Biochem. 1975; 53(8):914-20. DOI: 10.1139/o75-126. View

Zlatkis A, ZAK B, Boyle A . A new method for the direct determination of serum cholesterol. J Lab Clin Med. 1953; 41(3):486-92. View

BLIX G, Lindberg E, ODIN L, Werner I . Studies on sialic acids. Acta Soc Med Ups. 1956; 61(1-2):1-25. View

Cook G, Heard D, Seaman G . A sialomucopeptide liberated by trypsin from the human erythrocyte. Nature. 1960; 188:1011-2. DOI: 10.1038/1881011a0. View

Langley O, AMBROSE E . ISOLATION OF A MUCOPEPTIDE FROM THE SURFACE OF EHRLICH ASCITES TUMOUR CELLS. Nature. 1964; 204:53-4. DOI: 10.1038/204053a0. View

PORTEOUS J, Clark B . THE ISOLATION AND CHARACTERIZATION OF SUBCELLULAR COMPONENTS OF THE EPITHELIAL CELLS OF RABBIT SMALL INTESTINE. Biochem J. 1965; 96:159-71. PMC: 1206917. DOI: 10.1042/bj0960159. View

LOWRY O, ROSEBROUGH N, FARR A, RANDALL R . Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193(1):265-75. View

Chang K, Bennett V, Cuatrecasas P . Membrane receptors as general markers for plasma membrane isolation procedures. The use of 125-I-labeled wheat germ agglutinin, insulin, and cholera toxin. J Biol Chem. 1975; 250(2):488-500. View

Rouser G, Siakotos A, Fleischer S . Quantitative analysis of phospholipids by thin-layer chromatography and phosphorus analysis of spots. Lipids. 1966; 1(1):85-6. DOI: 10.1007/BF02668129. View

10.

Critchley D, Wyke J, Hynes R . Cell surface and metabolic labelling of the proteins of normal and transformed chicken cells. Biochim Biophys Acta. 1976; 436(2):335-52. DOI: 10.1016/0005-2736(76)90198-x. View

11.

Warley A, Cook G . The isolation and characterization of plasma membranes from normal and leukaemic cells of mice. Biochim Biophys Acta. 1973; 323(1):55-68. DOI: 10.1016/0005-2736(73)90431-8. View

12.

Gahmberg C, Simons K . Isolation of plasma membrane fragments from BHK21 cells. Acta Pathol Microbiol Scand B Microbiol Immunol. 1970; 78(2):176-82. DOI: 10.1111/j.1699-0463.1970.tb04284.x. View

13.

Kopaczyk K, Perdue J, Green D . The relation of structural and catalytic protein in the mitochondrial electron transfer chain. Arch Biochem Biophys. 1966; 115(1):215-25. DOI: 10.1016/s0003-9861(66)81060-3. View

14.

Perdue J, Sneider J . The isolation and characterization of the plasma membrane from chick embryo fibroblasts. Biochim Biophys Acta. 1970; 196(2):125-40. DOI: 10.1016/0005-2736(70)90001-5. View

15.

Van Lenten L, ASHWELL G . Studies on the chemical and enzymatic modification of glycoproteins. A general method for the tritiation of sialic acid-containing glycoproteins. J Biol Chem. 1971; 246(6):1889-94. View

16.

Fairbanks G, Steck T, Wallach D . Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971; 10(13):2606-17. DOI: 10.1021/bi00789a030. View

17.

Renkonen O, Gahmberg C, Simons K, Kaariainen L . The lipids of the plasma membranes and endoplasmic reticulum from cultured baby hamster kidney cells (BHK21). Biochim Biophys Acta. 1972; 255(1):66-78. DOI: 10.1016/0005-2736(72)90008-9. View

18.

Novogrodsky A, KATCHALSKI E . Membrane site modified on induction of the transformation of lymphocytes by periodate. Proc Natl Acad Sci U S A. 1972; 69(11):3207-10. PMC: 389737. DOI: 10.1073/pnas.69.11.3207. View

19.

Liao T, Gallop P, Blumenfeld O . Modification of sialyl residues of sialoglycoprotein(s) of the human erythrocyte surface. J Biol Chem. 1973; 248(23):8247-53. View

20.

Lloyd C, Cook G . On the mechanism of the increased aggregation by neuraminidase of 16C malignant rat dermal fibroblasts in vitro. J Cell Sci. 1974; 15(3):575-90. DOI: 10.1242/jcs.15.3.575. View