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小鼠L细胞干扰素的纯化。具有相关细胞生长抑制活性的基本纯品制剂。

Purification of mouse L cell interferon. Essentially pure preparations with associated cell growth inhibitory activity.

作者信息

Iwakura Y, Yonehara S, Kawade Y

出版信息

J Biol Chem. 1978 Jul 25;253(14):5074-9.

PMID:670178
Abstract

Mouse L cell interferon induced by Newcastle disease virus was purified by a combination of salt precipitation, ion exchange chromatography, gel filtration, and affinity chromatography using anti-interferon anti-body. The preparation was labeled with 125I at a later step of purification, which served as an index of protein concentration. Two main species of interferon molecules differing in molecular weight were separated from each other, and the final preparations obtained were shown to be essentially pure by polyacrylamide gel electrophoresis in the absence as well as in the presence of sodium dodecyl sulfate. The highest specific activities obtained were 2.6 X 10(9) and 7.3 X 10(8) international units/mg of protein (determined by 125I radioactivity) for the 40,000- and 24,000-dalton species, respectively. The preparations at different steps of purification, including those of the highest purity, were tested for the anti-cell growth activity. Their activities were found to be indistinguishable from each other when compared at the same antiviral doses, indicating that the anti-cell growth activity is carried by the interferon molecules themselves.

摘要

用盐析、离子交换层析、凝胶过滤以及使用抗干扰素抗体的亲和层析相结合的方法,纯化了新城疫病毒诱导产生的小鼠L细胞干扰素。在纯化的后期步骤中,用125I对该制剂进行标记,以此作为蛋白质浓度的指标。分离出了分子量不同的两种主要干扰素分子,通过在有无十二烷基硫酸钠存在的情况下进行聚丙烯酰胺凝胶电泳,结果表明最终获得的制剂基本纯净。对于分子量为40,000和24,000道尔顿的两种干扰素,所获得的最高比活性分别为2.6×10(9)和7.3×10(8)国际单位/毫克蛋白质(通过125I放射性测定)。对纯化不同阶段的制剂,包括纯度最高的制剂,进行了抗细胞生长活性测试。当在相同抗病毒剂量下进行比较时,发现它们的活性彼此无差异,这表明抗细胞生长活性由干扰素分子本身携带。

相似文献

1
Purification of mouse L cell interferon. Essentially pure preparations with associated cell growth inhibitory activity.小鼠L细胞干扰素的纯化。具有相关细胞生长抑制活性的基本纯品制剂。
J Biol Chem. 1978 Jul 25;253(14):5074-9.
2
Heterogeneity of purified mouse interferons.纯化小鼠干扰素的异质性。
J Biol Chem. 1975 Jun 10;250(11):4139-44.
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Interferon and cell division. VII. Inhibitory effect of highly purified interferon preparations on the multiplication of leukemia L 1210 cells.干扰素与细胞分裂。VII. 高度纯化的干扰素制剂对白血病L 1210细胞增殖的抑制作用。
Proc Soc Exp Biol Med. 1973 Jan;142(1):7-10. doi: 10.3181/00379727-142-36945.
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Purification of human lymphoblastoid interferon by a simple procedure with high yields.通过简单步骤高产率纯化人淋巴母细胞干扰素。
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Human lymphoblastoid interferon. Large scale production and partial purification.人淋巴母细胞干扰素。大规模生产与部分纯化。
J Biol Chem. 1977 Oct 10;252(19):6585-7.
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Preparative electrophoresis of isotopically labeled L-cell interferons.同位素标记的L细胞干扰素的制备电泳
Appl Microbiol. 1971 Jun;21(6):1067-71. doi: 10.1128/am.21.6.1067-1071.1971.
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SDS-polyacrylamide gel electrophoresis of purified human leucocyte interferon and the antiviral and anticellular activities of the different interferon species.纯化的人白细胞干扰素的SDS-聚丙烯酰胺凝胶电泳及不同干扰素种类的抗病毒和抗细胞活性
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Purification of mouse interferon by sequential affinity chromatography on poly(U)--and antibody--agarose columns.通过在聚(U)琼脂糖柱和抗体琼脂糖柱上进行连续亲和层析纯化小鼠干扰素。
Nature. 1978 Feb 16;271(5646):622-5. doi: 10.1038/271622a0.
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Purification of two components of mouse L cell interferon: electrophoretic demonstration of interferon proteins.小鼠L细胞干扰素两种成分的纯化:干扰素蛋白的电泳显示
J Gen Virol. 1976 Nov;33(2):225-36. doi: 10.1099/0022-1317-33-2-225.
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Purification and partial characterization of human lymphoblast interferon.人淋巴母细胞干扰素的纯化及部分特性鉴定
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引用本文的文献

1
Interferon potentiation occurs at a pretranscriptive stage.干扰素增强作用发生在转录前阶段。
Infect Immun. 1983 Jan;39(1):159-63. doi: 10.1128/iai.39.1.159-163.1983.
2
Ultrastructural localization of interferon receptors on the surfaces of cultured cells and erythrocytes.培养细胞和红细胞表面干扰素受体的超微结构定位
Infect Immun. 1982 May;36(2):811-21. doi: 10.1128/iai.36.2.811-821.1982.
3
Effect of interferon on chemiluminescence and hydroxyl radical production in murine macrophages stimulated by PMA.干扰素对佛波酯刺激的小鼠巨噬细胞化学发光及羟基自由基产生的影响。
Immunology. 1985 Nov;56(3):533-41.
4
Male sterility of transgenic mice carrying exogenous mouse interferon-beta gene under the control of the metallothionein enhancer-promoter.在金属硫蛋白增强子-启动子控制下携带外源小鼠干扰素-β基因的转基因小鼠的雄性不育。
EMBO J. 1988 Dec 1;7(12):3757-62. doi: 10.1002/j.1460-2075.1988.tb03259.x.
5
Regulation of cell proliferation and differentiation by interferons.干扰素对细胞增殖和分化的调控。
Biochem J. 1985 Mar 1;226(2):345-60. doi: 10.1042/bj2260345.
6
Purification and partial characterization of human lymphoblast interferon.人淋巴母细胞干扰素的纯化及部分特性鉴定
Proc Natl Acad Sci U S A. 1979 Nov;76(11):5601-5. doi: 10.1073/pnas.76.11.5601.
7
Effect of cholera toxin on the antiviral and anticellular activities of human leukocyte interferon.霍乱毒素对人白细胞干扰素抗病毒和抗细胞活性的影响。
Infect Immun. 1979 Oct;26(1):235-9. doi: 10.1128/iai.26.1.235-239.1979.