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用合成肽产生的白细胞介素-1抗体:人外周血单核细胞受刺激后白细胞介素-1外部位点的鉴定及合成分析

Antibodies to interleukin-1 raised with synthetic peptides: identification of external sites and analysis of interleukin-1 synthesis in stimulated human peripheral blood monocytes.

作者信息

Bomford R, Abdulla E, Hughes-Jenkins C, Simpkin D, Schmidt J

机构信息

Department of Experimental Immunobiology, Wellcome Research Laboratories, Beckenham, Kent, U.K.

出版信息

Immunology. 1987 Dec;62(4):543-9.

Abstract

Rabbit antibodies against peptides corresponding to amino acids 1-18, 45-65 and 71-90 of mature human interleukin-1 beta (IL-1 beta) precipitated 125I-labelled IL-1 beta, showing that these sites are accessible to antibody and located externally. Immunoprecipitation of 35S-methionine-labelled LPS-stimulated human peripheral blood monocytes followed by SDS-PAGE revealed the expected major bands of molecular weights 35,000 and 17,500. The 35,000 protein was found in the cell lysate and extracellularly in the medium, but the 17,500 protein was exclusively in the medium. A previously undescribed 31,000 band was also detected in the medium. These results are most simply explained by the hypothesis that the 35,000 IL-1 beta precursor is released from the cell and processed extracellularly to the 17,500 mature form. The 31,000 molecule may represent a processing intermediate.

摘要

针对成熟人白细胞介素-1β(IL-1β)氨基酸1-18、45-65和71-90对应肽段的兔抗体沉淀了125I标记的IL-1β,表明这些位点可被抗体识别且位于外部。对35S-甲硫氨酸标记的经脂多糖刺激的人外周血单核细胞进行免疫沉淀,随后进行SDS-PAGE,结果显示出预期的分子量为35,000和17,500的主要条带。分子量为35,000的蛋白质存在于细胞裂解物中以及培养基的细胞外,但分子量为17,500的蛋白质仅存在于培养基中。在培养基中还检测到一条先前未描述的分子量为31,000的条带。这些结果最简单的解释是基于这样的假设:分子量为35,000的IL-1β前体从细胞中释放出来,并在细胞外加工成分子量为17,500的成熟形式。分子量为31,000的分子可能代表加工中间体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b9f7/1454147/791f4d30fe7c/immunology00165-0036-a.jpg

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