Characterization of a soluble ternary complex formed between human interferon-β-1a and its receptor chains

被引：26

作者：

Arduini, RM ^{[1
]}

Strauch, KL ^{[1
]}

Runkel, LA ^{[1
]}

Carlson, MM ^{[1
]}

Hronowski, XP ^{[1
]}

Foley, SF ^{[1
]}

Young, CN ^{[1
]}

Cheng, WJ ^{[1
]}

Hochman, PS ^{[1
]}

Baker, DP ^{[1
]}

机构：

[1] Biogen Inc, Cambridge, MA 02142 USA

来源：

PROTEIN SCIENCE | 1999年 / 8卷 / 09期

关键词：

IFNAR1; IFNAR2; interferon-beta-1a; interferon receptor complexes;

D O I：

10.1110/ps.8.9.1867

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 extracellular portions of the chains that comprise the human type I interferon receptor, IFNAR1 and IFNAR2, have been expressed and purified as recombinant soluble His-tagged proteins, and their interactions with each other and with human interferon-beta-1a (IFN-beta-1a) were studied by gel filtration and by cross-linking. By gel filtration, no stable binary complexes between IFN-beta-1a and IFNAR1, or between IFNAR1 and IFNAR2 were detected. However, a stable binary complex formed between IFN-beta-1a and IFNAR2. Analysis of binary complex formation using various molar excesses of IFN-beta-1a and IFNAR2 indicated that the complex had a 1:1 stoichiometry, and reducing SDS-PAGE of the binary complex treated with the cross-linking reagent dissucinimidyl glutarate (DSG) indicated that the major cross-linked species had an apparent M-r consistent with the sum of its two individual components. Gel filtration of a mixture of IFNAR1 and the LFN-beta-1a/IFNAR2 complex indicated that the three proteins formed a stable ternary complex. Analysis of ternary complex formation using various molar excesses of IFNAR1 and the IFN-beta-1a/IFNAR2 complex indicated that the ternary complex had a 1:1:1 stoichiometry, and reducing SDS-PAGE of the ternary complex treated with DSG indicated that the major cross-linked species had an apparent M-r consistent with the sum of its three individual components. We conclude that the ternary complex farms by the sequential association of IFN-beta-1a with IFNAR2, followed by the association of LFNAR1 with the preformed binary complex. The ability to produce the IFN-beta-1a/IFNAR2 and IFN-beta-1a/IFNAR1/IFNAR2 complexes make them attractive candidates for X-ray crystallography studies aimed at determining the molecular interactions between IFN-beta-1a and its receptor.

引用

页码：1867 / 1877

页数：11

共 63 条

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