Ramanujam K S, Seetharam S, Seetharam B
Department of Medicine, Medical College of Wisconsin, Milwaukee 53226.
Biochim Biophys Acta. 1993 Mar 14;1146(2):243-6. doi: 10.1016/0005-2736(93)90362-4.
Intrinsic factor-cobalamin receptor (IFCR) activity in visceral yolk sac and placental membranes is regulated during pregnancy in rats. While the IFCR activity declined in the visceral yolk sac membranes by 15-fold, it rose nearly 20-fold in the placental membranes from fourteen to nineteen days of gestation. The visceral yolk sac membranes revealed a 230 kDa protein that co-migrated with pure rat renal IFCR. This 230 kDa band was also identified as IFCR in both the membranes by immunoblotting with anti-serum to rat renal IFCR. Immunoprecipitation of 35S labeled proteins obtained from in vitro translation using visceral yolk sac mRNA from 14-day pregnant rats, yielded on SDS-PAGE a single band of 220 kDa, while those obtained from 19-day pregnant rats did not. The binding of intrinsic factor-cyano[57Co]cobalamin complex to the visceral yolk sac membranes was inhibited by preincubation of these membranes with anti-serum to rat IFCR but not with anti-serum to rat asialoglycoprotein receptor or mannose or mannan or N-acetylglucosamine. Based on these results, we suggest that the IFCR activity, protein expression and mRNA levels in fetal membranes are regulated during pregnancy and may play an important role in the maternal-fetal transfer of cobalamin.
大鼠孕期内脏卵黄囊和胎盘膜中的内因子 - 钴胺素受体(IFCR)活性受到调节。在内脏卵黄囊膜中,IFCR活性下降了15倍,而在妊娠14至19天的胎盘膜中,其活性上升了近20倍。内脏卵黄囊膜显示出一种230 kDa的蛋白质,它与纯大鼠肾IFCR迁移位置相同。通过用抗大鼠肾IFCR血清进行免疫印迹,在这两种膜中也将这条230 kDa的条带鉴定为IFCR。对来自14天妊娠大鼠的内脏卵黄囊mRNA进行体外翻译获得的35S标记蛋白进行免疫沉淀,在SDS - PAGE上产生了一条220 kDa的单一条带,而来自19天妊娠大鼠的则没有。用抗大鼠IFCR血清预孵育这些膜可抑制内因子 - 氰基[57Co]钴胺素复合物与内脏卵黄囊膜的结合,但用抗大鼠去唾液酸糖蛋白受体血清、甘露糖、甘露聚糖或N - 乙酰葡糖胺预孵育则不能。基于这些结果,我们认为胎膜中的IFCR活性、蛋白质表达和mRNA水平在孕期受到调节,并且可能在钴胺素的母胎转运中起重要作用。