INHIBITION OF INTERLEUKIN-2/P55 RECEPTOR SUBUNIT INTERACTION BY COMPLEMENTARY PEPTIDES
INHIBITION OF INTERLEUKIN-2/P55 RECEPTOR SUBUNIT INTERACTION BY COMPLEMENTARY PEPTIDES
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DOI：
10.1006/abbi.1995.1201
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发表时间：
1995-04-01
影响因子：
3.9
通讯作者：
ISETTA, AM
中科院分区：
文献类型：
--
作者：
FASSINA, G;CASSANI, G;ISETTA, AM
对白介素a(IL-2)序列的互补肽进行了模拟天然受体和抑制IL-2/P55受体亚单位相互作用的能力测试。以线性和多聚体的形式合成了与IL-2序列15-27和40-54具有水路互补性的多肽，然后首先用固相结合试验表征了它们与IL-2相互作用的能力。多聚体互补多肽与生物素化的IL-2之间的结合是特异的、可饱和的，并且被线性和多聚体互补多肽抑制，以微摩尔范围内的解离常数为特征的饱和相互作用也发生在微滴板上的IL-2与生物素化的线性和多聚体互补多肽之间，与IL-2靶序列相对应的多肽能够干扰这种相互作用，以及全长IL-2。肽的识别是序列依赖的，因为互补肽序列或IL-2靶序列的扰乱消除了结合，固相载体上固定后的多聚体互补肽也被证明是有效的，直接从粗品混合物中亲和纯化与靶部位相对应的重组IL-2或IL-2片段，产量高，回收率高。IL-2序列15-27的互补多肽，而不是IL-2序列40-54的线性或多聚体形式的互补多肽，即使具有不同的效力，也会在体外干扰IL-2/P55受体亚单位的相互作用，从而提示该IL-2位点在受体识别中可能起作用。(C)1995年学术出版社。
Complementary peptides to interleukin-a (IL-2) sequences important for receptor binding were tested for their ability to mimic natural receptors and act as inhibitors of the IL-2/p55 receptor subunit interaction. Peptides hydropathically complementary to IL-2 sequences 15-27 and 40-54 were synthesized in a linear and in a multimeric form and then characterized first by solid-phase binding assays for their ability to interact with IL-2. Binding between the multimeric complementary peptides and biotinylated IL-2 was specific, saturable, and inhibited by linear as well as multimeric complementary peptides, Saturable interactions, characterized by dissociation constants in the micromolar range, occurred also between IL-2 immobilized on microtiter plates and biotinylated linear and multimeric complementary peptides, Peptides corresponding to the IL-2 target sequences were able to interfere with this interaction, as well as full-length IL-2. Peptide recognition was sequence dependent, since scrambling of complementary peptide sequences or IL-2 target peptide sequences abolished binding, Multimeric complementary peptides after immobilization on solid supports proved useful also for affinity purifications of recombinant IL-2 or IL-2 fragments corresponding to the target sites, directly from crude mixtures, in high yield and with high recovery. Complementary peptides to IL-2 sequence 15-27, but not to IL-2 sequence 40-54, in the linear or in the multimeric form, even if with different potency, interfered with the IL-2/p55 receptor subunit interaction in vitro, thus suggesting a possible role of this IL-2 site in receptor recognition. (C) 1995 Academic Press,Inc.