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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  • [31] CRYSTAL-STRUCTURE OF THE CYTOKINE INTERLEUKIN-1-BETA
    PRIESTLE, JP
    SCHAR, HP
    GRUTTER, MG
    [J]. EMBO JOURNAL, 1988, 7 (02) : 339 - 343
  • [32] LOW AND HIGH-AFFINITY CELLULAR RECEPTORS FOR INTERLEUKIN-2 - IMPLICATIONS FOR THE LEVEL OF TAC ANTIGEN
    ROBB, RJ
    GREENE, WC
    RUSK, CM
    [J]. JOURNAL OF EXPERIMENTAL MEDICINE, 1984, 160 (04) : 1126 - 1146
  • [33] RORDORFADAM C, 1989, DRUG EXP CLIN RES, V15, P355
  • [34] SCHAR HP, 1987, J BIOL CHEM, V262, P13724
  • [35] SECKINGER P, 1987, J IMMUNOL, V139, P1546
  • [36] SEGAL DM, 1977, J IMMUNOL, V118, P1338
  • [37] INTERLEUKIN-1 SIGNALING OCCURS EXCLUSIVELY VIA THE TYPE-I RECEPTOR
    SIMS, JE
    GAYLE, MA
    SLACK, JL
    ALDERSON, MR
    BIRD, TA
    GIRI, JG
    COLOTTA, F
    RE, F
    MANTOVANI, A
    SHANEBECK, K
    GRABSTEIN, KH
    DOWER, SK
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (13) : 6155 - 6159
  • [38] CDNA EXPRESSION CLONING OF THE IL-1 RECEPTOR, A MEMBER OF THE IMMUNOGLOBULIN SUPERFAMILY
    SIMS, JE
    MARCH, CJ
    COSMAN, D
    WIDMER, MB
    MACDONALD, HR
    MCMAHAN, CJ
    GRUBIN, CE
    WIGNALL, JM
    JACKSON, JL
    CALL, SM
    FRIEND, D
    ALPERT, AR
    GILLIS, S
    URDAL, DL
    DOWER, SK
    [J]. SCIENCE, 1988, 241 (4865) : 585 - 589
  • [39] SIMS JE, 1990, YEAR IMMUNOLOGY 1989, V6, P112
  • [40] SLEATH PR, 1990, J BIOL CHEM, V265, P14526