Di Trapani G, Orosco C, Perkins A, Clarke F
Division of Science and Technology, Griffith University, Nathan, Queensland, Australia.
Hum Reprod. 1991 Mar;6(3):450-7. doi: 10.1093/oxfordjournals.humrep.a137357.
Protocols are described for the isolation from human placental extracts of a preparation which is active in the rosette inhibition assay by inducing an increased rosette inhibition titre. In this respect the preparation mimics the effects of pregnancy sera, an ability ascribed to the presence in these sera of a so-called 'early pregnancy factor' (EPF). In addition to this activity in the rosette inhibition assay, the preparation was also shown to modulate the expression of cell surface immunoglobulin on peripheral blood mononuclear cells. The polypeptide composition of the preparation was relatively simple, as revealed by SDS-PAGE. There was a major 12 kd polypeptide previously isolated in a rosette inhibition titre-active preparation from ovine placental extracts, and a small amount of 68 kd polypeptide. N-Terminal amino acid sequence analyses obtained after blotting onto polyvinylidene difluoride membranes identified the latter polypeptide as serum albumin and the major 12 kd polypeptide as human thioredoxin. Gel permeation analysis partially resolved activity expression away from the 12 kd polypeptide as activity expression was also found to be associated with low mol. wt material. It is concluded that EPF activity expression in pregnancy sera involves molecules related to and associated with thioredoxin.
本文描述了从人胎盘提取物中分离一种制剂的方法，该制剂在玫瑰花结抑制试验中具有活性，可诱导玫瑰花结抑制效价升高。在这方面，该制剂模拟了妊娠血清的作用，这种能力归因于这些血清中存在一种所谓的“早孕因子”（EPF）。除了在玫瑰花结抑制试验中的这种活性外，该制剂还被证明可调节外周血单个核细胞上细胞表面免疫球蛋白的表达。如SDS-PAGE所示，该制剂的多肽组成相对简单。有一种主要的12kd多肽，先前从绵羊胎盘提取物中分离出的一种具有玫瑰花结抑制效价活性的制剂中含有这种多肽，还有少量68kd多肽。将其印迹到聚偏二氟乙烯膜上后进行的N端氨基酸序列分析确定，后一种多肽为血清白蛋白，主要的12kd多肽为人硫氧还蛋白。凝胶渗透分析部分地将活性表达与12kd多肽分离开来，因为活性表达也被发现与低分子量物质有关。得出的结论是，妊娠血清中EPF活性的表达涉及与硫氧还蛋白相关和关联的分子。
