Purification and characterization of a uterine retinol-binding protein in the bitch.
Purification and characterization of a uterine retinol-binding protein in the bitch.
复制标题
母犬子宫视黄醇结合蛋白的纯化和表征。
DOI：
10.1042/bj3110407
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Cotton,M
中科院分区：
文献类型：
--
作者：
Buhi,WC;Alvarez,IM;Shille,VM;Thatcher,MJ;Harney,JP;Cotton,M
通过离子交换和凝胶过滤色谱法纯化了一种主要的犬子宫内膜分泌蛋白（cP 6，23，000-M（r）），并通过二维凝胶电泳进行了表征。抗[人视黄醇结合蛋白（hRBP）]血清在免疫印迹分析中鉴定了cP 6，并从培养基中免疫沉淀了cP 6。这种主要蛋白质也显示出结合[3 H]视黄醇。确定N-末端和内部氨基酸序列，并与先前鉴定的蛋白质、RNA或DNA序列进行比较。N-末端分析表明，cP 6与血清视黄醇结合蛋白（RBP）具有高度的同一性和相似性，而内部序列分析表明，与大鼠雄激素依赖性附睾蛋白和β-乳球蛋白具有很强的相似性。然而，氨基酸分析表明，这些蛋白质和cP 6在总氨基酸含量和某些选定的氨基酸之间存在显着差异。免疫组化分析显示RBP仅在子宫腔上皮中染色。这些研究表明，母狗子宫内膜分泌一个蛋白质家族（cP 6），其中一些结合[3 H]视黄醇，与RBP家族免疫相关，并且具有与RBP、β-乳球蛋白和脂质运载蛋白家族其他成员高度相似的N-末端和内部序列。这个蛋白质家族在早期发育中可能很重要，为发育中的胚胎提供视黄醇或衍生物。
A major canine endometrial secreted protein (cP6, 23,000-M(r)) was purified by ion-exchange and gel-filtration chromatography and characterized by two-dimensional gel electrophoresis. Anti-[human retinol-binding protein (hRBP)] serum identified cP6 on immunoblot analysis and immunoprecipitated cP6 from culture medium. This major protein was also shown to bind [3H]retinol. N-terminal and internal amino acid sequences were determined and compared with previously identified protein, RNA, or DNA sequences. N-terminal analysis revealed that cP6 had high identity and similarity to serum retinol-binding proteins (RBPs), while internal sequence analysis showed a strong similarity to rat androgen-dependent epididymal protein and beta-lactoglobulins. Amino acid analysis, however, showed significant differences between these proteins and cP6 in both total amino acid content and certain selected amino acids. Immunohistochemical analysis showed staining for RBP only in the uterine luminal epithelium. These studies suggest that bitch endometrium secretes a family of proteins (cP6), some of which bind [3H]retinol, are immunologically related to the RBP family, and have N-terminal and internal sequences with a high similarity to RBP, beta-lactoglobulins and other members of the lipocalin family. This family of proteins may be important in early development for supplying retinol or derivatives to the developing embryo.