Abstract
Murine extra-embryonic endodermal cell lines derived from either teratocarcinomas or mouse embryos contain a cytoskeletal protein (Endo A) of Mr = 55,000. Endo A was immunoprecipitated from [35S]methionine-labeled lysates of three parietal endodermal cell lines, A presumptive visceral endodermal cell line, and a fetal hepatoma cell line, but not from fibroblasts, myoblasts, erythroleukemic cells, neuroblastoma cells, keratinocytes, or embryonal carcinoma cells. Embryonal carcinoma cells induced to differentiate by exposure to retinoic acid synthesized increased amounts of Endo A approximately 48 h after exposure to the inducer. Two-dimensional gel analysis of immunoprecipitated samples confirmed that Endo A is distinct from vimentin and murine keratinocyte proteins recognized by two different keratin antisera. Comparison by two-dimensional gel electrophoresis of immunoprecipitated Endo A labeled with either [35S]methionine or [32P]orthophosphate indicated that the multiple forms of Endo A resolved by isoelectric focusing were due, at least in part, to phosphorylation. Serine was identified as the phosphorylated amino acid. Endo A was the only major antigenic protein found in a parietal endodermal cell line which was recognized by a monoclonal antibody prepared by other investigators against trophoblast cytoskeletons. The results indicate that Endo A, like the previously described Endo B protein, is distinct from other cytoskeletal proteins and will be useful as a marker of the differentiation of murine embryonal carcinoma cells to extra-embryonic endoderm.
Published Version
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