Folate binding protein from kidney brush border membranes contains components characteristic of a glycoinositol phospholipid anchor.
Folate binding protein from kidney brush border membranes contains components characteristic of a glycoinositol phospholipid anchor.
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来自肾刷状缘膜的叶酸结合蛋白含有糖肌醇磷脂锚的特征成分。
DOI：
10.1021/bi00127a027
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发表时间：
1992
期刊：
影响因子：
2.9
通讯作者：
Rosenberry,TL
中科院分区：
文献类型：
--
作者：
Lee,HC;Shoda,R;Krall,JA;Foster,JD;Selhub,J;Rosenberry,TL
修订稿于 1991 年 12 月 4 日收到 摘要：许多细胞表面蛋白已被证明通过与成熟蛋白 C 末端的酰胺键共价连接的糖肌醇磷脂 (GPL) 锚定在质膜上。我们应用了几个标准来确定大鼠肾刷状缘膜中的叶酸结合蛋白 (FBP) 含有 GPL 锚。分离刷状缘膜并用 [3H] 叶酸标记，通过与纯化的细菌磷脂酰肌醇特异性磷脂酶 C (PIPLC) 一起孵育，FBP 和 [3H] 叶酸复合物被释放到上清液中，但与纯化的细菌磷脂酰胆碱特异性磷脂酶 C 一起孵育则不会。粗提物中的 FBP-[3H] 叶酸复合物以及通过配体定向亲和层析纯化 FBP 后与 Triton 相互作用Xl 14 胶束和预先与 PIPLC 一起孵育可防止这种去污剂相互作用。在十二烷基硫酸钠中进行聚丙烯酰胺凝胶电泳后，发现 GPL 锚定特征的各个残基与 FBP 共价关联。其中包括葡萄糖胺和乙醇胺（通过还原甲基化进行放射性标记，并通过氨基酸分析仪上的色谱法进行鉴定）和肌醇磷酸盐（通过使用抗 CRD 抗血清的蛋白质印迹法推断）。仅在 FBP 被 PIPLC（GPL 锚定的可靠诊断）裂解后，该抗血清才产生阳性免疫染色。还讨论了生物膜中 GPL 锚定的 FBP 与生物体液中可溶性 FBP 之间的关系。 整合膜蛋白通常通过一个或多个跨膜片段中的疏水氨基酸残基与磷脂双层膜的疏水相保持紧密接触 (Sabatini et al., 1982)。这些蛋白质需要去污剂来溶解并结合提取物中的去污剂。然而，已经很明显的是
Revised Manuscript Received December 4, 1991 abstract: A number of cell surface proteins have been shown to be anchored to the plasmamembrane by a covalently attached glycoinositol phospholipid (GPL) in amide linkage to the C-terminus of the mature protein. We applied several criteria to establish that folate binding protein (FBP) in brush border membranes of rat kidney contains a GPL anchor. Brush border membranes were isolated and labeled with [3H] folate, and the complex of FBP and [3H] folate was shown to be released to the supernatant by incubation with purified bacterial phosphatidylinositol-specific phospholipase C (PIPLC) but not by incubation with a purified bacterial phosphatidylcholine-specific phospholipase C. The FBP-[3H] folate complex both in crude extracts and after FBP purification by ligand-directed affinity chromatography interacted withTriton Xl 14 micelles, and prior incubation with PIPLC prevented this detergent interaction. Individual residues characteristic of GPL anchors were found to be covalently associated with FBP following polyacrylamide gel electrophoresis in sodium dodecyl sulfate. These included glucosamine and ethanolamine, which were radiolabeled by reductive methylation and identified by chromatography on an amino acid analyzer, and inositol phosphate, which was inferred by Western blotting with an anti-CRD antisera. This antisera gave positive immu-nostaining only after FBP had been cleaved by PIPLC, a reliable diagnostic of a GPL anchor. The relationship between GPL-anchored FBP in biological membranes and soluble FBP in biological fluids also is discussed.Integral membrane proteins generally maintainintimate contact with the hydrophobic phase of phospholipid bilayer membranes through hydrophobic amino acid residues in one or more transmembrane segments (Sabatini et al., 1982). These proteins require detergents for solubilization and bind detergents in extracts. However, it has become apparent that
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影响因子：
56.9
作者：
FERGUSON, MAJ;HOMANS, SW;RADEMACHER, TW
通讯作者：
RADEMACHER, TW
人红细胞乙酰胆碱酯酶 C 末端糖脂膜结合域中胺成分的鉴定。
影响因子：
2.9
作者：
Haas,R;Brandt,PT;Knight,J;Rosenberry,TL
通讯作者：
Rosenberry,TL
人胎盘中 Mg2 依赖性蛋白酶的鉴定，该蛋白酶将疏水性叶酸结合蛋白裂解为亲水性形式。
DOI：
--
发表时间：
1989
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Antony,AC;Verma,RS;Unune,AR;LaRosa,JA
通讯作者：
LaRosa,JA
激素响应性肾质膜组分的制备和表征。
影响因子：
4.8
作者：
S. Marx;S. Fedak;G. Aurbach
通讯作者：
G. Aurbach
DOI：
10.1042/bj2440001
发表时间：
1987-05
期刊：
The Biochemical journal
影响因子：
--
作者：
Martin G. Low
通讯作者：
Martin G. Low