Properties of biologically active messenger RNA from human placenta. Cell-free synthesis of two immunoreactive forms of placental lactogen.
In order to understand better the regulation of human placental proteins the activity of placental lactogen messenger RNA has been examined. Total RNA was extracted from normal term placentas and purified by chromatography on oligo(dT)-cellulose. The poly(A)-containing fraction stimulated amino acid incorporation 5- to 10-fold in wheat germ cell-free extracts, and immunoprecipitation of the translation products with antiserum directed against human placental lactogen (hPL) suggests that about 2% of the peptides contain hPL determinants. Analysis of the material precipitated with hPL antiserum by electrophoresis on sodium dodecyl sulfate-polyacrylamide gels revealed two major species, one co-migrating with hPL and the other migrating slightly slower than hPL. On DEAE-cellulose chromatography the former material eluted close to authentic hPL while the latter material eluted at higher ionic strength than hPL, indicating a difference in net charge of these two species. Tryptic peptide analysis of the large material and authentic hPL shows marked similarities in the primary structure of these two proteins. The slower migrating peptide has an apparent molecular weight about 3000 larger than hPL and thus may represent a precursor molecule. Both cell-free products could be competed out of immunoprecipitates by a large excess of authentic hPL, confirming their immunologic similarities. Centrifugation of the placental poly(A)-containing RNA through aqueous glycerol gradients indicates that the hPL mRNA sediments at about 14 S.
D049148:Aminoacylation; D000937:Antigen-Antibody Reactions; D002474:Cell-Free System; D005260:Female; D010446:Peptide Fragments / Q000032:analysis; D010920:Placenta / Q000378:metabolism*; D010928:Placental Lactogen / Q000096:biosynthesis*; D011247:Pregnancy; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism; D012639:Seeds / Q000378:metabolism; D014908:Triticum / Q000378:metabolism
D010446:Peptide Fragments; D012333:RNA, Messenger; D010928:Placental Lactogen