ISOLATION OF THE NON-GLYCOSYLATED PROTEINS OF DESMOSOMES AND IMMUNOLOCALIZATION OF A 3RD PLAQUE PROTEIN - DESMOPLAKIN-III
ISOLATION OF THE NON-GLYCOSYLATED PROTEINS OF DESMOSOMES AND IMMUNOLOCALIZATION OF A 3RD PLAQUE PROTEIN - DESMOPLAKIN-III
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DOI：
10.1073/pnas.82.3.810
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发表时间：
1985-01-01
影响因子：
11.1
通讯作者：
STEINBERG, MS
中科院分区：
文献类型：
--
作者：
GORBSKY, G;COHEN, SM;STEINBERG, MS
The cytoplasmic plaque of the spot desmosome or macula adherens mediates the attachment of bundles of intermediate filaments to the plasma membrane. A fraction was isolated from a bovine epidermal desmosome preparation that is highly enriched in the non-glycosylated desmosomal proteins. Plastic-embedded and thin-sectioned high-speed pellets of this fraction reveal closely packed filaments that resemble plaque regions of the low pH whole desmosome preparation from which they are derived. NaDodSO4 [sodium dodecyl sulfate]/polyacrylamide gel electrophoresis reveals 4 major, non-glycosylated proteins of 240, 210, 81 and 77 kDa [kilodalton]. The 240- and 210-KDa proteins (desmoplakins I and II) are closely related, whereas the 81- and 77-kDa proteins are unique. This is shown both immunologically and by 1-dimensional proteolytic peptide mapping. Monospecific, polyclonal rabbit antibodies were prepared against the 81-kDa protein and used, in conjunction with protein A-complexed colloidal gold particles (PAG), to immunolocalize this antigen on ultrathin sections of bovine muzzle epidermis. On antibody-labeled sections, PAG particles were associated principally with the desmosomal cytoplasmic plaque. Sections exposed to preimmune serum showed little or no labeling. The 81-kDa protein, like the 240/210-kDa protein family, is one of the major components of the desmosomal plaque. It is designated as desmoplakin III. The location of the 77-kDa protein remains to be definitively established.
