Isolation and partial characterization of a pregnancy-associated glycoprotein family from the goat placenta

Abstract

Antigen(s) immunologically related to pregnancy-associated glycoproteins (PAGs) have previously been detected in the serum of pregnant goats. In this work, we describe a partial characterization of a family of PAGs isolated from the placenta of the goat. The procedure, monitored by RIA, included extraction of proteins at neutral pH, acidic, and ammonium sulfate precipitations; and gel filtration and ion exchange chromatographies. Immunoreactivity, initially located in the acidic supernatant and in the 40- 80% ammonium sulfate fractions, was equally apportioned between the 0.04 and 0.08 M NaCl DEAE fractions. After further purification of both DEAE fractions, the preparations were subjected to one- and two-dimensional electrophoresis, and individual polypeptides were analyzed by amino acid sequencing. Three PAGs, which differed in amino acid sequence and apparent molecular masses (62, 59, and 55 kDa), were detected, each containing several isoforms with different pls: caprine (c) PAG62 (pI: 5.1, 4.8), cPAG59 (pI: 6.2, 5.9, 5.6), and cPAG55 (pI: 5.3, 5.1, 4.9). These proteins had high sequence identities to each other and to PAGs purified from other species. Each had two putative N-glycosylation sites within the 27 amino terminal residues sequenced. This work demonstrates that PAGs are present in goat placenta and that multiple forms are expressed.