Characterization of bovine conceptus proteins produced during the peri- and postattachment periods of early pregnancy.
Characterization of bovine conceptus proteins produced during the peri- and postattachment periods of early pregnancy.
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早期妊娠围产期和附着后时期产生的牛概念蛋白的特征。
DOI：
10.1095/biolreprod38.3.703
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发表时间：
1988
影响因子：
3.6
通讯作者：
Gillespie,BE
中科院分区：
文献类型：
--
作者：
Godkin,JD;LifseyJr,BJ;Gillespie,BE
在胎盘形成的围栏期和贴壁后期（分别为第17-24天和第26-38天），将从子宫中取出的牛孕体在含L-[3 H]亮氨酸的改良最低必需培养基中培养，以表征释放到培养基中的蛋白质的体外合成。蛋白质生产模式进行了分析，通过二维聚丙烯酰胺凝胶电泳，然后通过干凝胶荧光。观察到四组低分子量酸性蛋白（LMWAP）在围栏和附着后期间合成。这些的数量和相对浓度随着发展而变化。一组（A）由3种主要和2种或2种以上次要等电物质（pI> 5.8-6.8）组成;这些是第17-22天观察到的主要合成蛋白质。A组的主要多肽在第38天检查的所有时间点均存在，并且在几种制备物中，在第29天之前（但之后）出现双峰（Mr为22，000和24，000）。通过免疫印迹分析证明来自第19天和第36天的孕体培养物的A组多肽与针对绵羊滋养层蛋白-1（oTP-1）产生的抗血清交叉反应。在第17天至第22天期间观察到第二组蛋白质（A1）和20，000 - 24，000 Mr范围内的单一蛋白质（B）。这些相对于A组为酸性，并且在第22天后未检测到。第四组（C）的LMWAP（Mr 14，000 - 18，000）在第21天左右首次观察到，并且似乎相对于A组增加至第29天。该组的一种蛋白质C3是第38天的主要LMWAP。在第21天和第24天之间首次观察到在30，000 - 70，000 Mrrange（E组）中产生许多蛋白质。其中三种-E7、E8和E9-是酸性的（pI = 5.0-5.8），表观分子量在30，000和45，000之间，在第29天左右和之后的时间成为主要的分泌产物。从第17天到至少第29天，存在非常大的（Mr> 200，000）酸性（pI < 4.5）蛋白，在第24天前后和之后观察到几个大的（Mr> 100，000）蛋白。A组蛋白质被称为牛滋养层蛋白-1（bTP-1），并涉及与母体识别妊娠相关的事件。本研究是第一个证明这些产品的持续生产超过第24天，并报告在早期postattachment期间观察到的牛孕体蛋白质合成的定性变化。
Bovine conceptuses removed from the uterus during the pen- and postattachment periods of placentation (Days 17–24 and 26–38, respectively) were cultured in a modified minimum essential medium in the presence of L-[3H] leucine to characterize in vitro synthesis of proteins released into the medium. Patterns of protein production were analyzed by two-dimensional polyacrylamide gel electrophoresis followed by fluorography of dried gels. Four groups of low molecular weight acidic proteins (LMWAP) were observed to be syn¬thesized during the pen- and postattachment periods. The number and relative concentration of these changed with development. One group (A) consisted of three major and two or more minor isoelectric species (pI ≅ 5.8–6.8); these were the major synthesized proteins observed from Days 17–22. The major polypeptides of Group A were present at all time points examined through Day 38 and, in several preparations, appeared as doublets (Mr≃ 22,000 and 24,000) through Day 29 but not thereafter. Group A polypeptides from Day 19 and 36 conceptus cultures were demonstrated by immunoblot analysis to cross-react with antiserum produced against ovine trophoblast protein-1 (oTP-1). A second group of proteins (A1) and a single protein (B) in the 20,000–24,000 Mr range were observed between Days 17 and 22. These were acidic relative to Group A and were not detected after Day 22. A fourth group (C) of LMWAP (Mr≅ 14,000–18,000) was first observed around Day 21 and appeared to increase relative to Group A through Day 29. One protein from this group, C3, was the predominant LMWAP at Day 38. Production of numerous proteins in the 30,000–70,000 Mrrange (Group E) was first observed between Days 21 and 24. Three of these—E7, , E8, and E9—were acidic (pI ≃ 5.0–5.8), with apparent molecular weights between 30,000 and 45,000, and became prominent secretory products around Day 29 and times thereafter. A very large (Mr> 200,000) acidic (pI < 4.5) protein was present from Day 17 through at least Day 29, and several large (Mr> 100,000) were observed around Day 24 and afterwards. Group A proteins have been referred to as bovine trophoblast protein-1 (bTP-1) and implicated in events associated with maternal recognition of pregnancy. The present study is the first to demonstrate continued production of these products beyond Day 24, and to report qualitative changes observed in bovine conceptus protein synthesis during the early postattachment period.