Preparation of a human lung purified plasma membrane fraction: confirmation by enzyme markers, electron microscopy, and histamine H1 receptor binding

The Journal of Membrane Biology
T B CasaleM Kaliner

Abstract

A simple and rapid method of isolating plasma membranes from human peripheral lung tissue is described. The method involves homogenization of tissue in 0.25 M sucrose-buffered medium followed by differential and sucrose density gradient centrifugation. Enzymatic and morphological characterization of the plasma membrane fraction revealed minimal contamination by nonplasma membrane fragments. The isolated plasma membranes showed an 18-fold purification of 5'-nucleotidase activity compared to the original homogenate. Electron-microscopic studies of the plasma membrane fraction revealed the presence of small membrane vesicles having a trilaminar membrane structure. To further examine the purity of the plasma membrane preparation, the binding of the H1 receptor antagonist, 3H pyrilamine, to the plasma membrane-enriched fraction was compared to the binding to crude membrane preparations. Both the plasma membrane-enriched fraction and the crude membrane preparation had similar Kd's for the histamine antagonist, but the plasma membrane-enriched fraction had a threefold greater binding capacity, reflecting the relative enrichment of plasma membranes of the preparation. Thus, a method has been developed for the isolation of plasma membra...Continue Reading

References

Sep 11, 1979·Life Sciences·J C Schwartz
Oct 1, 1979·The Journal of Allergy and Clinical Immunology·L SteelM Kaliner
Jul 1, 1979·Journal of Molecular and Cellular Cardiology·C Y KwanE E Daniel
Sep 1, 1972·The Journal of Experimental Medicine·M KalinerK F Austen
Sep 9, 1967·Biochimica Et Biophysica Acta·R ColemanJ N Hawthorne
Aug 1, 1972·Biochemistry·R R WeihingA H Phillips

Citations

Feb 1, 1990·American Journal of Respiratory Cell and Molecular Biology·J P Joad
Dec 1, 1990·American Journal of Respiratory Cell and Molecular Biology·M CluzelL M Lichtenstein

Related Concepts

Plasma Membrane
Cytochrome c Oxidase Subunit VIa
Cytological Techniques
Lactate Dehydrogenase
Lung
Cell Surface Proteins
Electron Microscopy
Cytochrome c Reductase
Nucleotidases
Receptors, Histamine H1

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