Characterization of Proteins Produced by Porcine Embryonic and Amniotic Tissues Using Two-Dimensional Electrophoresis

以雙向電泳分析豬的胎兒和羊膜所分泌的蛋白質之特性

劉光輝(Kaung-Huei Liu) ； George A. Baumbach

《中華民國獸醫學會雜誌》 22卷3期 (1996/06) Pp. 153-162

https://doi.org/10.7009/JTCSVS.199606.0153

摘要

將妊娠第21天豬的胎兒及妊娠第21、55和112天豬的羊膜分別培養在含35S-甲硫胺酸或3H-白胺酸的培養液24小時。利用雙向電泳配合螢光顯像技術，分析早期胎兒和羊膜所分泌的蛋白質之特性。妊娠第21天的胎兒能分泌一些蛋白質；它們在雙向電泳膠上分佈之情形與血漿中血清蛋白非常類似。除此之外，以免疫沉澱技術，一個分子量為22,000的蛋白質被證實為源自胎兒的“維生素A 結合蛋白質”(RBP)。妊娠第21至55天的羊膜主要分泌一組名為B2的蛋白質。B2蛋白已被部份純化出；其可能是由兩個多胜肽所組成。於分娩前期，羊膜分泌蛋白質的能力大為下降。羊水內含有血清蛋白、RBP和B2蛋白。本篇報告的結果將有助於未來分析與比較在不同病理狀況下，對胎兒和羊膜蛋白質分泌的能力之影響。部份純化及分析B2蛋白之特性，提供未來純化並鑑定其生理功能必需的資料。

關鍵字

胎兒 ； 羊膜 ； 維生素A

並列摘要

Porcine embryos on day 21 of pregnancy, and amniotic membranes on day 21, 55 and 112 of pregnancy were cultured 24 h in the minimum essential medium in the presence of [35S] methionine or [3H] leucine. Proteins synthesized and released into the medium during culture were analyzed by two-dimensional polyacrylamide gel electrophoresis and fluorography. The day 21 embryos produced an array of polypeptides which exhibited electrophoretic mobilities and spatial arrangement on gels nearly identical to serum proteins (transferring, α-fetoprotein, α1-antitrypsin and fibrinogen) in the plasma. In addition, a protein with a molecular weight of 22,000 was identified as the embryonic retinol-binding protein (RBP) which was determined by immunoprecipitation with an anti-RBP serum. A group of proteins, designated as the B2 complex, were the predominant polypeptide products of amnion between days 21 and 55 of pregnancy. The B2 complex which probably existed as a dimer was partially purified. Prior to parturition, capacity for release of marcromolecule products by amnion was decreased. Serum protein-like proteins, RBP and B2 complex were present in the amniotic fluid. These data should help analyze the changes in patterns of proteins produced by embryo and amnion of pig evoked by pathological conditions. Partial purification and characterization of the B2 complex set the stage for future investigations toward identification of the function of the major amniotic protein products.

並列關鍵字

Embryo ； Amnion ； Vitamim A