Isolation of Desmosomes from the Epidermis of Xenopus laevis and Immunochemical Characterization of the Xenopus Desmosomal Cadherins

Cell Structure and Function

Hisato Shida²  Rie Ohga²  Mariko Shida¹ 

[1] Department of Food and Nutrition, Yamanashi Gakuin Junior College;Laboratory of Molecular Cell Biology, Faculty of Medicine, Yamanashi University

关键词: isolation;    desmosome;    immunochemical characterization;    desmosomal cadherin;    Xenopus laevis;    epidermis;   

DOI  :  10.1247/csf.29.17

学科分类：分子生物学，细胞生物学和基因

来源: Japan Society for Cell Biology

【 摘 要 】

References(45)Cited-By(1)Here we report a new method of isolating epidermal desmosomes from Xenopus laevis, and a major constituent of desmosomes designated as Xenopus desmogleins (XDsg). Isolation of desmosomes from Xenopus laevis epidermis was carried out by a two step-incubation with different concentrations of NP-40. After discontinuous sucrose gradient centrifugation at 30,000 g for 60 min, a pure desmosomal fraction was obtained at 30%/40% interface. In the SDS-PAGE of isolated desmosomes, at least 12 bands (XDB1 to XDB12) were observed over a 75 kD region. Among them, three bands (XDB3, XDB7, XDB8; estimated MW 175, 124, and 112 kD respectively) were recognized as glycoproteins based on ConA binding. Monospecific polyclonal antibody against XDB3 cross-reacted with bovine Dsgs and vis-a-vis anti-bovine Dsgs with XDB3. By contrast, monospecific antibody against bovine Dsc a/b did not cross-react with either XDB7 or XDB8. Heterogeneous molecular constituents of desmosomal adhesion molecule, which have been observed among different bovine tissues, were confirmed in a phylogenetically different animal, Xenopus laevis. Combined results with other evidence could suggest an alternative system for desmosome-mediated cell adhesion.

【 授权许可】

Unknown