ISOLATION OF EPIDERMAL DESMOSOMES
Open Access
- 1 November 1974
- journal article
- research article
- Published by Rockefeller University Press in The Journal of cell biology
- Vol. 63 (2), 515-523
- https://doi.org/10.1083/jcb.63.2.515
Abstract
A method is reported for the isolation of desmosomes in a high yield and of a purity suitable for biochemical analysis. The procedure utilizes the selective solubilizing action of citric acid-sodium citrate (CASC) buffer, pH 2.6, on the non-cornified layers of cow nose epidermis, followed by discontinuous sucrose density gradient centrifugation. Electron microscopy with both thin sections of pellets and unfixed spread preparations reveals that after centrifugation, desmosomes are located mainly at the 55–60% sucrose interface. In the desmosome preparation thus obtained, the characteristic desmosome structure is well preserved, showing the midline, unit membranes, and dense plaques. Furthermore, removal of the epidermal filament bundles by the solubilizing action of CASC buffer has revealed a finely filamentous layer on the cytoplasmic surface of the plaques. The dimensions, location, and appearance of this layer correspond with those of the "connecting component" which has been previously suggested as being responsible for the attachment of epidermal filament bundles to the desmosome.Keywords
This publication has 13 references indexed in Scilit:
- PLASMA MEMBRANES OF MAMMALIAN CELLSThe Journal of cell biology, 1973
- Membrane ultrastructure at mammalian intercellular junctionsProgress in Biophysics and Molecular Biology, 1973
- LANTHANUM STAINING OF THE SURFACE COAT OF CELLSThe Journal of cell biology, 1971
- Ultrastructure of intercellular junctions in adult and developing cardiac muscleThe American Journal of Cardiology, 1970
- THE ULTRASTRUCTURE OF THE CAT MYOCARDIUMThe Journal of cell biology, 1969
- Urea extraction of Z bands, intercalated disks, and desmosomesJournal of Ultrastructure Research, 1968
- An electron-microscopic study of the junctional and regular desmosomes in normal human epidermia.1968
- The Membrane Protein of Horny Cells**From the Department of Dermatology, Boston University School of Medicine, Boston University Medical Center, Boston, Mass.Journal of Investigative Dermatology, 1966
- FINE STRUCTURE OF DESMOSOMES, HEMIDESMOSOMES, AND AN ADEPIDERMAL GLOBULAR LAYER IN DEVELOPING NEWT EPIDERMISThe Journal of cell biology, 1966
- CELL JUNCTIONS IN AMPHIBIAN SKINThe Journal of cell biology, 1965