Biosynthesis and turnover of a 34-kDa protein growth factor in human cytotrophoblasts.
Recently we isolated a new protein growth factor of 34 kDa from synctial membranes of human placenta. In its polypeptide molecular mass, antigenic structure, receptor binding specificity and partial amino acid sequence, it is unlike several known growth factors, hormones and other proteins. Here we report studies on its biosynthesis and turnover in cultured cytotrophoblasts from term human placenta. Expression of the 34-kDa protein in these cells was studied by immunoprecipitation and Western blot analyses using a highly specific antibody. The experiments have produced the following results. a) Immunostaining and Western blot analyses have demonstrated the presence of immunoreactive 34-kDa protein in isolated cytotrophoblasts. The protein is present in both freshly isolated cells and in cells that have fused in culture to form multinuclear syncytiotrophoblasts. b) Trophoblastic biosynthesis of the protein has been demonstrated by in vitro translation of cellular mRNA and by metabolic labelling experiments with intact cells. c) Pulse-chase experiments show that biosynthesis of the protein does not involve any detectable precursors of higher or lower molecular mass. d) Studies on turnover indicate that the synthesized protein is unusually stable with a half-life of 50-70 h.
Roy-Choudhury
S
S
Department of Biochemistry, University of Pennsylvania School of Medicine, Philadelphia 19104.
Sen-Majumdar
A
A
Murthy
U
U
Mishra
V S
VS
Kliman
H J
HJ
Nestler
J E
JE
Strauss
J F
JF
Das
M
M
D002478:Cells, Cultured; D005260:Female; D006133:Growth Substances / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy; D014176:Protein Biosynthesis; D011506:Proteins / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D014327:Trophoblasts / Q000378:metabolism*
D006133:Growth Substances; D011506:Proteins; D012333:RNA, Messenger