Protein synthesis and secretion in human decidua of early pregnancy.
Proteins synthesized and secreted by first trimester decidua in primary culture were identified. Explants were cultured for 24 h, in RPMI-1640 or Dulbecco's Modified Eagles Medium containing the radioactive amino acid 35S-methionine or 3H-proline. Electron microscopy of explants before and after 24 h of culture demonstrated the relative purity of the decidua, maintenance of cell integrity, and ultrastructural features indicative of active protein synthesis and secretion. Proteins synthesized and secreted by the explants into the medium were analyzed by fluorography of one-dimensional polyacrylamide gels in the presence of sodium dodecyl sulfate. By comparison of radiolabeled proteins from four women, eight 35S-methionine-labeled bands of 78, 70, 60, 50, 43, 34, 25, and 23 kDa were identified as common decidual peptides. A comparison of autoradiographs of the medium from decidual cultures to decidual cell homogenates showed that seven of these peptides were enriched in the culture medium. When labeled peptides from fibroblast cultures were compared to labeled proteins from decidual cultures each of the common decidual peptides (except the 70 kDa protein) occurred only in the decidual culture medium. Comparison of 3H-proline and 35S-methionine-labeled decidual proteins revealed that the 78, 70, 60, 50, and 34 kDa proteins were of similar fluorographic intensity when labeled with the two different amino acids. The 43, 25, and 23 kDa proteins appeared to contain more methionine, and proteins at 36, 20, 13, and 12 kDa were proline-rich, but contained less methionine. The seven decidual explant-specific, 35S-methionine-labeled secreted proteins were concentrated and purified by preparative gel electrophoresis, and antisera were generated to four of the putative decidual secretory proteins.
D000918:Antibody Specificity; D003656:Decidua / Q000378:metabolism* / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007120:Immunochemistry; D066298:In Vitro Techniques; D008854:Microscopy, Electron; D011247:Pregnancy / Q000378:metabolism*; D011261:Pregnancy Trimester, First; D014176:Protein Biosynthesis; D011506:Proteins / Q000276:immunology / Q000378:metabolism
D011506:Proteins