Purification of desmoglein II: a method for the preparation and fractionation of desmosomal components.
We have developed rapid and efficient methods for the isolation of desmosomes and the fractionation of their components. These methods involve the use of 6 M guanidine HCl to isolate the desmosomes from bovine epidermis, followed by hydroxyapatite column chromatography in the presence of SDS to fractionate the desmosomal components. All of the desmoplakins and desmogleins were purified at least partially by these procedures, and desmoglein II was purified to apparent homogeneity. We expect these procedures to facilitate a detailed biochemical analysis of the molecular components of desmosomes. In addition, these methods may be applicable to the purification of other plasma membrane domains involved in cell adhesion.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D002458:Cell Fractionation / Q000379:methods; D002845:Chromatography / Q000379:methods; D003598:Cytoskeletal Proteins; D051301:Desmoglein 2; D051182:Desmogleins; D051180:Desmoplakins; D003896:Desmosomes / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D005340:Fibrinogen / Q000031:analogs & derivatives* / Q000378:metabolism; D008565:Membrane Proteins / Q000302:isolation & purification*; D008854:Microscopy, Electron
D000596:Amino Acids; D003598:Cytoskeletal Proteins; D051301:Desmoglein 2; D051182:Desmogleins; D051180:Desmoplakins; D008565:Membrane Proteins; C050126:aglycofibrinogen; D005340:Fibrinogen