A novel glycoprotein of the aspartic proteinase gene family expressed in bovine placental trophectoderm.
The pregnancy associated glycoproteins (PAG 1) that appear in the maternal serum of cattle and sheep soon after implantation are apparently inactive members of the aspartic proteinase family. Here we describe the isolation of a highly abundant cDNA (PAG 2 cDNA) that represents a second member of this gene family which is structurally related to bovine PAG 1, ovine PAG 1, and pepsin (58%, 58%, and 51% amino acid sequence identity, respectively). The bovine PAG 2 cDNA was identified in two ways. First, the bovine placental library was screened under relatively nonstringent conditions with an ovine PAG 1 cDNA. The second fortuitous approach employed immunoscreening with an antiserum raised against a partially purified factor that competed with bovine LH for binding to the LH receptor on the CL of the ovary. The full-length cDNA (1258 bp) codes for a polypeptide of 376 amino acids. Bovine PAG 2, unlike bovine PAG 1, has a catalytic center with a consensus sequence of amino acids. Its mRNA is expressed in fetal placenta but not in other fetal organs, and is localized to both the mononucleate and binucleate cells of the trophectoderm, whereas PAG 1 is expressed only in binucleate cells. PAG 2 is synthesized by placental explants as a 70-kDa glycoprotein that is processed to several smaller molecules. Western blot analysis of culture media developed with epitope-selected antibodies to PAG 2 reveals several bands ranging in apparent M(r) from 31,000-70,000, which correspond in size to the polypeptides present in the preparation used for immunization.(ABSTRACT TRUNCATED AT 250 WORDS)
Xie
S
S
Department of Animal Sciences, University of Missouri-Columbia 65211.
Low
B G
BG
Nagel
R J
RJ
Beckers
J F
JF
Roberts
R M
RM
D000595:Amino Acid Sequence; D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D015152:Blotting, Northern; D015153:Blotting, Western; D002417:Cattle; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D000939:Epitopes / Q000032:analysis; D005260:Female; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D010920:Placenta / Q000737:chemistry / Q000201:enzymology* / Q000378:metabolism; D011233:Precipitin Tests; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D012756:Sheep; D014327:Trophoblasts / Q000166:cytology / Q000201:enzymology*
D018076:DNA, Complementary; D000939:Epitopes; D011257:Pregnancy Proteins; D016282:Aspartic Acid Endopeptidases