Pig interleukin 1. Purification of two immunologically different leukocyte proteins that cause cartilage resorption, lymphocyte activation, and fever
Pig interleukin 1. Purification of two immunologically different leukocyte proteins that cause cartilage resorption, lymphocyte activation, and fever
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猪白细胞介素 1. 纯化两种免疫学不同的白细胞蛋白，导致软骨吸收、淋巴细胞活化和发热
DOI：
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发表时间：
1985
影响因子：
15.3
通讯作者：
Y. Townsend
中科院分区：
文献类型：
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作者：
J. Saklatvala;S. Sarsfield;Y. Townsend
从刀豆蛋白A刺激的猪黄皮毛细胞培养上清液中纯化出两种形式的白介素1(IL-1)，经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法测定，两种蛋白的等电点均为21,000，但其中一种为软骨吸收蛋白，其等电点为5(IL-1/5)，另一种为等电点8.3(IL-1/8)。经初步凝胶过滤和聚焦层析分离后，用羟基磷灰石和离子交换剂Mono-S和Mono-Q层析纯化IL-1/5，用Monos-S在pH 4.0和pH 6.4的条件下层析纯化IL-1/8，用软骨蛋白多糖释放法检测纯化情况，两种蛋白的最终比活力均约为白细胞培养上清液的10(5)倍。用200份L血培养上清液，可产生约15微克和50微克IL-1/8。IL-1/8可促进小鼠胸腺细胞增殖，刺激滑膜成纤维细胞产生前列腺素E和潜伏性胶原酶。IL-1/5先前已被证明具有所有这些活性。每种IL-1在兔体内均可产生抗血清。每种抗血清在软骨生物测定中抑制其各自的IL-1，并对其进行Western blotting染色。抗血清对另一种IL-1既无抑制作用，也无染色作用。我们的结论是，猪白细胞产生两种不同的免疫形式的IL-1，具有相同的MR，显示相同的生物活性范围，但等电点不同。
Two forms of interleukin 1 (IL-1) were purified to homogeneity from the culture supernatants of pig buffy coat leukocytes stimulated with concanavalin A. The two proteins had identical Mr of 21,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but one, which had previously been purified as a cartilage-resorbing protein, had pI 5 (IL- 1/5) and the other, pI 8.3 (IL-1/8). After initial gel filtration and separation by chromatofocusing IL-1/5 was purified by chromatography on hydroxyapatite and the ion exchangers, Mono S and Mono Q; IL-1/8 was purified by chromatography at pH 4.0 and pH 6.4 on Mono S. Purification was monitored by a cartilage proteoglycan release assay and both proteins had a final specific activity approximately 10(5) times that of the leukocyte culture medium. Medium conditioned by cells from 200 L of blood yielded approximately 15 micrograms of IL-1/5 and 50 micrograms IL-1/8. IL-1/8 augmented mouse thymocyte proliferation, stimulated synovial fibroblasts to produce prostaglandin E and latent collagenase, and was pyrogenic upon intracerebroventricular injection into rabbits. IL-1/5 has previously been shown to possess all these activities. An antiserum to each IL-1 was raised in rabbits. Each antiserum inhibited its respective IL-1 in a cartilage bioassay and stained it upon Western blotting. Neither antiserum inhibited or stained the other IL-1. We conclude that pig leukocytes make two immunologically distinct forms of IL-1 that have identical Mr, demonstrate the same range of biological activity, but differ in isoelectric point.
DOI：
10.1073/pnas.81.24.7907
发表时间：
1984-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
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作者：
AURON, PE;WEBB, AC;DINARELLO, CA
通讯作者：
DINARELLO, CA