The pregnancy-associated glycoproteins (PAGs) are secretory products synthesized by the outer epithelial cell layer (chorion) of the placentas of various ungulate species. The amino acid sequences of eight PAGs have been inferred from cloned cDNA of cattle and sheep, as well as of the non-ruminant pig and horse. We compare the PAG sequences and present results of the three-dimensional models of boPAG-1 and ovPAG-1 that were constructed on the basis of the crystal structures of homologous porcine pepsin and bovine chymosin using a rule-based comparative modelling approach. Further, we compare peptide binding subsites defined by interactions with pepstatin and a decapeptide inhibitor (CH-66) modelled on the basis of crystal structures of other aspartic proteinases. We have extended our analysis of the peptide binding subsites to the other PAG molecules of known sequence by aligning the PAG sequences to the structural template derived from the pepsin family and by making use of the three-dimensional models of the boPAG-1 and ovPAG-1. The residues that are likely to affect peptide binding in the boPAG-1, ovPAG-1 and other PAG molecules have been identified. Sequence comparisons reveal that all PAG molecules may have evolved from a pepsin-like progenitor molecule with the equine PAG most closely related to the pepsins. The presence of substitutions at the S₁ and other subsites relative to pepsin make it unlikely that either bovine, ovine or the porcine PAG-1 have catalytic activity. Only two of the eight PAGs examined (porcine PAG-2 and equine PAG-1) retain features of active aspartic proteinases with pepsin-like activity. Our results indicate that in the PAGs so far characterized the peptide binding specificities differ significantly from each other and from pepsin, despite their high sequence identities. Analysis of the various peptide binding subsites demonstrates why both bovine and ovine PAG-1 are capable of binding pepstatin. The strong negative charge in the binding cleft of boPAG-1 and ovPAG-1 indicates a preference for lysine- or arginine-rich peptides. PAGs represent a family where the possible peptide binding function may be retained through their binding specificities, but where the catalytic activity may be lost in some cases, such as the boPAG-1, ovPAG-1 and the poPAG-1.

Reproduction Nutrition Development, formerly Annales de Biologie Animale Biochimie Biophysique

The effect of prolonged hyperprolactinaemia on the secretion of LH, progesterone and oestradiol, and its relationship to the maintenance of pregnancy was examined in pigs. Twelve crossbred, pregnant gilts were injected i.m. with 1.5 mg haloperidol kg−1 body weight (n = 6) or vehicle (n = 6) once a day from day 60 to day 66 of pregnancy. Blood samples were collected at 08:00, 12:00, 16:00, 20:00, 24:00 h from day 60 to day 67 and every 15 min for 4 h (08:00–12:00 h) h) on days 60, 63 and 66. Plasma concentrations of prolactin were higher (P < 0.001) in haloperidol-treated gilts than in control gilts (121.3 ± 4.3 ng ml−1 and 13.6 ± 0.4 ng ml−1, respectively). Hyperprolactinaemia completely inhibited the pulsatile secretion of LH and diminished (P < 0.001) basal peripheral concentrations of LH (hyperprolactinaemia, 0.3 ± 0.04 ng ml−1 and control, 0.6 ± 0.005 ng ml−1). Despite the inhibition of LH release in hyperprolactinaemic gilts, plasma concentrations of progesterone were higher (P < 0.001) than in the control group (20.8 ± 0.6 and 12.6 ± 0.2 ng ml−1, respectively). Oestradiol concentrations were not different between groups, although oestradiol tended to be higher in hyperprolactinaemic gilts than in the control group throughout the sampling period (29.1 ± 1.9 versus 23.7 ± 1.6 pg ml−1, respectively). Abortion did not occur in any of the gilts. These results are the first to demonstrate that induced hyperprolactinaemia during the second half of pregnancy (days 60–66) will drastically suppress the major porcine luteotrophin but not affect pregnancy maintenance in pigs. It is possible that prolactin has a more important function in the luteotrophic complex as an additional protectant associated with the regulatory mechanism of late pregnancy maintenance than was previously reported.