Rat clusterin isolated from primary Sertoli cell-enriched culture medium is sulfated glycoprotein-2 (SGP-2).
Rat clusterin isolated from primary Sertoli cell-enriched culture medium is sulfated glycoprotein-2 (SGP-2).
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DOI：
10.1016/s0006-291x(88)81099-4
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发表时间：
1988-08
影响因子：
3.1
通讯作者：
C. Cheng;C. L. Chen;Z. Feng;A. Marshall;C. Bardin
中科院分区：
文献类型：
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作者：
C. Cheng;C. L. Chen;Z. Feng;A. Marshall;C. Bardin
白细胞蝶呤是一种最初从公羊睾丸网液中分离的糖蛋白，是由具有不同NH 2-末端氨基酸序列的单体组成的二聚体。鉴于其在生精上皮细胞间相互作用中的可能作用，我们试图在大鼠中鉴定这样的蛋白质。使用开发的羊蛋白的生物测定，大鼠clusterin纯化至表观均匀性，通过HPLC从初级支持细胞富集培养基。该蛋白也是由Mr 43，000（α）和Mr 35，000（β）单体组成的异二聚体。氨基酸序列分析表明，α亚基的氨基酸序列为NH 2-SLMPLSHYGPLSFHNMFQPFFDMIHQAQQA，β亚基的氨基酸序列为NH 2-EQEFSDNELQELSTQGSRYVNKEIQNAVQG。这两个亚基与从睾丸网液中分离的公羊聚集素的相应亚基具有显著的相似性。用抗大鼠聚集蛋白（α）亚基的抗体，从大鼠睾丸λ gt 11 cDNA文库中克隆了一个cDNA克隆。来自分离的大鼠丛生蛋白cDNA的氨基酸序列和NH 2-末端氨基酸序列的分析表明，大鼠丛生蛋白是相同的支持细胞糖蛋白以前指定的硫酸化糖蛋白-2。
Clusterin, a glycoprotein originally isolated from ram rete testis fluid, is a dimer composed of monomers with non-identical NH2-terminal amino acid sequences. In view of its possible role in cell-cell interactions in the seminiferous epithelium, we sought to identify such a protein in the rat. Using the bioassay developed for the ovine protein, rat clusterin was purified to apparent homogeneity by HPLC from primary Sertoli cell-enriched culture media. This protein is also a heterodimer consisting of monomers of Mr 43,000 (α) and Mr 35,000 (β). NH2-Terminal amino acid sequence analysis indicated that the (α) subunit has a sequence of NH2-SLMPLSHYGPLSFHNMFQPFFDMIHQAQQA and the β subunit, NH2-EQEFSDNELQELSTQGSRYVNKEIQNAVQG. These two subunits show marked similarity with the corresponding subunits of ram clusterin isolated from rete testis fluid. Using an antibody against the (α) subunit of rat clusterin, a cDNA clone was isolated from a rat testicular lambda gt11 cDNA library. Analyses of the amino acid sequence derived from the isolated rat clusterin cDNA and of the NH2-terminal amino acid sequences indicate that rat clusterin is identical to a Sertoli cell glycoprotein previously designated sulfated glycoprotein-2.