A new trophoblast-derived growth factor from human placenta: purification and receptor identification.
This paper describes the identification and characterization of a new peptide growth factor. The peptide was isolated from trophoblastic brush border membranes of human placenta. The purified preparation was homogeneous and consisted of a single polypeptide of Mr 34 000 with a pI of about 6.0. This peptide stimulated DNA replication in cultured fibroblasts. The following association was seen between activity and protein: During DEAE-cellulose chromatography, both the 34-kilodalton (kDa) protein and the mitogenic activity displayed identical binding and salt dependence of elution. Nondenaturing electrophoresis at pH 8.3 revealed a comigration of the 34-kDa protein and the DNA replication stimulatory activity. Identical electrophoretic mobilities were displayed for both activity and protein at pH 7.0. These results demonstrate that the preparation is homogeneous and show that growth factor activity is intrinsic to the 34-kDa polypeptide. Binding of the 125I-labeled 34-kDa mitogen to target fibroblastic cells was specific; i.e., nanomolar concentrations of the unlabeled 34-kDa protein competed effectively with the labeled protein, whereas a variety of well-characterized growth factors and hormones were unable to compete even at micromolar levels. Thus the 34-kDa protein interacts with target cells through highly specific surface receptors. Chemical cross-linking techniques were used to investigate the identity of the receptor for the 34-kDa mitogen. Cross-linking of fibroblastic cells containing bound 125I-labeled 34-kDa protein generated a radiolabeled complex of 86 kDa in all four cell types examined.(ABSTRACT TRUNCATED AT 250 WORDS)
D000818:Animals; D001667:Binding, Competitive; D002478:Cells, Cultured; D004261:DNA Replication / Q000187:drug effects; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D051379:Mice; D008871:Microvilli / Q000378:metabolism; D008970:Molecular Weight; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D011817:Rabbits; D013045:Species Specificity; D014327:Trophoblasts / Q000378:metabolism*
D011257:Pregnancy Proteins; C048517:trophoblast-derived growth factor