A MUTATIONAL ANALYSIS OF RECEPTOR-BINDING SITES OF INTERLEUKIN-1-BETA - DIFFERENCES IN BINDING OF HUMAN INTERLEUKIN-1-BETA MUTEINS TO HUMAN AND MOUSE RECEPTORS

被引:31
作者
GRUTTER, MG [1 ]
VANOOSTRUM, J [1 ]
PRIESTLE, JP [1 ]
EDELMANN, E [1 ]
JOSS, U [1 ]
FEIGE, U [1 ]
VOSBECK, K [1 ]
SCHMITZ, A [1 ]
机构
[1] CIBA GEIGY AG, DEPT INFLAMMAT BONE & ALLERGY, DIV PHARMACEUT, CH-4002 BASEL, SWITZERLAND
来源
PROTEIN ENGINEERING | 1994年 / 7卷 / 05期
关键词
INTERLEUKIN-1-BETA; MUTATIONAL MAPPING; RECEPTOR BINDING; SPECIES DIFFERENCE;
D O I
10.1093/protein/7.5.663
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The 3-D crystal structure of interleukin-1 beta (IL-1 beta) has been used to define its receptor binding surface by mutational analysis. The surface of IL-1 beta was probed by site-directed mutagenesis. A total of 27 different IL-1 beta muteins were constructed, purified and analyzed. Receptor binding measurements on mouse and human cell lines were performed to identify receptor affinities. IL-1 beta muteins with modified receptor affinity were evaluated for structural integrity by CD spectroscopy or X-ray crystallography. Changes in six surface loops, as well as in the C- and N-termini, yielded muteins with lower binding affinities. Two muteins with intact binding affinities showed 10- to 100-fold reduced biological activity. The surface region involved in receptor binding constitutes a discontinuous area of similar to 1000 Angstrom(2) formed by discontinuous polypeptide chain stretches. Based on these results, a subdivision into two distinct local areas is proposed. Differences in receptor binding affinities for human and mouse receptors have been observed for some muteins, but not for wild-type IL-1 beta. This is the first time a difference in binding affinity of IL-1 beta muteins to human and mouse receptors has been demonstrated.
引用
收藏
页码:663 / 671
页数:9
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