Godkin J D, Lifsey B J, Gillespie B E
Department of Animal Science, University of Tennessee, Knoxville 37901-1071.
Biol Reprod. 1988 Apr;38(3):703-11. doi: 10.1095/biolreprod38.3.703.
Bovine conceptuses removed from the uterus during the peri- 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 synthesized during the peri- and postattachment periods. The number and relative concentrations of these changed with development. One group (A) consisted of three major and two or more minor isoelectric species (pI approximately equal to 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 approximately equal to 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 approximately equal to 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.(ABSTRACT TRUNCATED AT 250 WORDS)
在胎盘形成的附着期及附着后期(分别为第17 - 24天和第26 - 38天)从子宫中取出的牛孕体,在添加L - [³H]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情况。通过二维聚丙烯酰胺凝胶电泳,随后对干燥凝胶进行荧光显影,分析蛋白质产生模式。在附着期及附着后期观察到四组低分子量酸性蛋白(LMWAP)被合成。这些蛋白的数量和相对浓度随发育而变化。一组(A)由三个主要和两个或更多次要等电点物种(pI约等于5.8 - 6.8)组成;这些是在第17 - 22天观察到的主要合成蛋白。A组的主要多肽在第38天之前的所有检测时间点都存在,并且在一些制剂中,直到第29天都表现为双峰(Mr约等于22,000和24,000),之后则没有。通过免疫印迹分析证明,来自第19天和第36天孕体培养物的A组多肽与针对羊滋养层蛋白 - 1(oTP - 1)产生的抗血清发生交叉反应。在第17天至第22天之间观察到第二组蛋白(A1)和一个分子量在20,000 - 24,000 Mr范围内的单一蛋白(B)。相对于A组,这些蛋白呈酸性,在第22天之后未检测到。第四组LMWAP(C组,Mr约等于14,000 - 18,000)在第21天左右首次观察到,并且相对于A组似乎在第29天之前有所增加。该组中的一种蛋白C3在第38天是主要的LMWAP。(摘要截于250字)