期刊
JOURNAL OF BIOLOGICAL CHEMISTRY
卷 279, 期 21, 页码 22773-22780出版社
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
DOI: 10.1074/jbc.M313660200
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Plakin family members envoplakin and periplakin have been shown to be part of the cornified cell envelope in terminally differentiating stratified squamous epithelia. In the present study, purified recombinant human envoplakin and periplakin were used to investigate their properties and interactions. We found that envoplakin was insoluble at physiological conditions in vitro, and co- assembly with periplakin was required for its solubility. Envoplakin and periplakin formed soluble complexes with equimolar stoichiometry. Chemical cross- linking revealed that the major soluble form of all periplakin constructs and of envoplakin/ periplakin rod domains was a dimer, although co- assembly of the full-length proteins resulted in formation of higher order oligomers. Electron microscopy of rotary- shadowed periplakin demonstrated thin flexible molecules with an average contour length of 88 nm for the rod- plus- tail fragment, and immunolabeling EM confirmed the molecule as a parallel, in- register, dimer. Both periplakin and envoplakin/ periplakin oligomers were able to bind synthetic lipid vesicles whose composition mimicked the cytoplasmic side of the plasma membrane of eukaryotic cells. This binding was dependent on anionic phospholipids and Ca2+. These findings raise the possibility that envoplakin and periplakin bind to the plasma membrane upon elevation of intracellular [ Ca2+] in differentiating keratinocytes, where they serve as a scaffold for cornified cell envelope assembly.
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