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将氚标记的前体掺入大鼠嗜碱性白血病细胞IgE受体中。

The incorporation of tritiated precursors into receptors for IgE of rat basophilic leukemia cells.

作者信息

Helm R M, Froese A

出版信息

Immunology. 1981 Apr;42(4):629-36.

Abstract

It has previously been shown that two receptors, H and R, having apparent mol. wt of 55,000 and 45,000 daltons, can be isolated from surface iodinated RBL cells. In addition, small amounts of a third molecule of 71,000 daltons (71 K) can now be detected in receptor preparations isolated by means of IgE-Sepharose. The SDS-PAGE patterns show that the three surface molecules were not significantly altered when cell solubilization was carried out in the presence of protease inhibitors. The same three molecules were also observed when cells were biosynthetically labelled with [3H]-fucose of [3H]-galactose. When cells were labelled with [3H]-leucine or [3H]-tyrosine, several additional molecules were observed in IgE-Sepharose purified preparations. When cells were exposed to free IgE- before solubilization, significant inhibition of binding to IgE-Sepharose could only be seen among molecules corresponding to H, R and 71 K. When receptors were isolated by means of DNP-IgE and anti-DNP-Sepharose, fewer molecules were found. One, which was frequently observed, had a mol. wt of 26,000 daltons. This molecule may be non-covalently associated with receptors for IgE.

摘要

先前的研究表明,可从经表面碘化的大鼠嗜碱性粒细胞白血病(RBL)细胞中分离出两种受体,即H受体和R受体,其表观分子量分别为55,000道尔顿和45,000道尔顿。此外,现在在通过IgE-琼脂糖凝胶分离的受体制剂中可检测到少量分子量为71,000道尔顿(71K)的第三种分子。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)图谱显示,当在蛋白酶抑制剂存在的情况下进行细胞溶解时,这三种表面分子没有明显改变。当用[3H]-岩藻糖或[3H]-半乳糖对细胞进行生物合成标记时,也观察到了相同的三种分子。当用[3H]-亮氨酸或[3H]-酪氨酸对细胞进行标记时,在IgE-琼脂糖凝胶纯化的制剂中观察到了几种其他分子。当细胞在溶解前暴露于游离IgE时,仅在与H、R和71K相对应的分子中观察到对与IgE-琼脂糖凝胶结合的显著抑制。当通过二硝基苯基(DNP)-IgE和抗DNP-琼脂糖凝胶分离受体时,发现的分子较少。经常观察到的一种分子的分子量为26,000道尔顿。该分子可能与IgE受体非共价结合。

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