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干扰素-α诱导的人78kD蛋白:纯化及其与小鼠Mx蛋白的同源性、单克隆抗体的制备以及干扰素-γ的增强作用

IFN-alpha induced human 78 kD protein: purification and homologies with the mouse Mx protein, production of monoclonal antibodies, and potentiation effect of IFN-gamma.

作者信息

Horisberger M A, Hochkeppel H K

机构信息

Ciba-Geigy Ltd., Pharmaceutical Research, Basel, Switzerland.

出版信息

J Interferon Res. 1987 Aug;7(4):331-43. doi: 10.1089/jir.1987.7.331.

Abstract

A 78-kD protein (p78) is induced in human cells in response to interferon (IFN). It appeared as a radioactive spot when newly synthesized proteins from IFN-treated human cells labeled with [35S]methionine were separated in a two-dimensional system and autoradiographed. p78 was induced by IFN-alpha in normal human fibroblasts, and in some, but not all, established human tumor cell lines. It has been purified to homogeneity from Namalwa cells induced by recombinant IFN-alpha. Mouse polyclonal and monoclonal antibodies specific to p78 have been produced which allowed its quantitative determination in a Western blot ELISA. Using this method it was also shown that although IFN-gamma was a poor inducer of p78, it markedly increased the effect of IFN-alpha on p78 induction and accumulation. It was also demonstrated that p78 and the protein Mx of influenza-resistant mice, which we purified and characterized earlier (Horisberger and Hochkeppel, J. Biol. Chem. 260, 1730-1733, 1985) share common properties such as size, pI, amino acid composition, antigenic determinant(s), and IFN inducibility.

摘要

一种78-kD蛋白(p78)在人类细胞中受干扰素(IFN)诱导而产生。当用[35S]甲硫氨酸标记的经IFN处理的人类细胞新合成的蛋白质在二维系统中分离并进行放射自显影时,它表现为一个放射性斑点。p78在正常人成纤维细胞以及一些(但不是全部)已建立的人类肿瘤细胞系中由IFN-α诱导产生。它已从重组IFN-α诱导的Namalwa细胞中纯化至同质。已制备出针对p78的小鼠多克隆和单克隆抗体,这使得可以在蛋白质印迹酶联免疫吸附测定中对其进行定量测定。使用这种方法还表明,尽管IFN-γ是p78的弱诱导剂,但它显著增强了IFN-α对p78诱导和积累的作用。还证明了p78与我们之前纯化和鉴定的抗流感小鼠的Mx蛋白(Horisberger和Hochkeppel,《生物化学杂志》260,1730 - 1733,1985)具有共同特性,如大小、pI、氨基酸组成、抗原决定簇以及IFN诱导性。

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