Differentiation-dependent changes in the solubility of a 195-kD protein in human epidermal keratinocytes.
We have prepared a monoclonal antibody, AE11, that recognizes specifically a 195-kD protein (pI 5.4) of human keratinocytes. This antigen constitutes approximately 0.01-0.1% of total protein in keratinocytes of skin, esophagus, and cornea, and is readily detectable in these cells by immunofluorescent staining and immunoblotting. However, it is barely detectable in MCF mammary carcinoma cells and HeLa cells, and is undetectable in nonepithelial cell types. Results from serial extraction experiments have shown that this protein exists in two distinct pools: a Tris-soluble, and a Tris-insoluble but urea- or SDS-soluble one. The distribution of the 195-kD protein between these two pools appears to be differentiation-related, since relatively undifferentiated cells selected by a low-calcium medium contain primarily the soluble form, while highly differentiated cells contain mainly the insoluble form. Data from immunofluorescent staining and trypsin-sensitivity experiments suggest that the soluble form is cytoplasmic, whereas the insoluble form is submembranously located at the cell periphery of upper, differentiated cells. The insoluble, cell peripheral form of the 195-kD antigen increases progressively during epidermal differentiation; its insolubility appears to be related to the formation of disulfide-bond(s). These results indicate that the 195-kD protein, which has recently been suggested to be involved in cornified envelope formation (Simon, M., and H. Green, 1985, Cell, 36:827-834), undergoes significant changes in its solubility characteristics and intracellular location during keratinocyte maturation.
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D002451:Cell Compartmentation; D002454:Cell Differentiation; D003593:Cytoplasm / Q000378:metabolism; D004817:Epidermis / Q000648:ultrastructure*; D005455:Fluorescent Antibody Technique; D006801:Humans; D008970:Molecular Weight; D011506:Proteins / Q000276:immunology / Q000378:metabolism*; D012995:Solubility
D000911:Antibodies, Monoclonal; D011506:Proteins