Consequences of IkappaB alpha hydroxylation by the factor inhibiting HIF (FIH)
Consequences of IkappaB alpha hydroxylation by the factor inhibiting HIF (FIH)
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DOI：
10.1016/j.febslet.2010.10.060
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发表时间：
2010-12-01
期刊：
影响因子：
3.5
通讯作者：
Komives, Elizabeth A.
中科院分区：
文献类型：
--
作者：
Devries, Ingrid L.;Hampton-Smith, Rachel J.;Komives, Elizabeth A.
抑制HIF-1(FIH-1)的因子使许多含有锚蛋白重复序列的蛋白羟化，包括I kappa Bα。人们普遍推测，I kappa Bα的羟化具有功能后果，但羟化的影响尚未得到证实。我们制备了羟化的I kappa Bα，并将其与未羟化的蛋白质进行了比较。尿素变性和酰胺H/D交换实验表明，羟化后的“折叠性”没有变化。结合核因子kappaB的表面等离子体共振测量表明，核因子kappaB结合的动力学或热力学没有差别。泛素非依赖性蛋白酶体降解实验表明，蛋白质的半衰期没有差别。因此，FIH-1对I-kappa Bα的羟基化似乎是无关紧要的，至少对于我们可以在体外测定的功能是不重要的。结构摘要：MINT-8051494：NF-kappa-B p65(uniprotkb：Q04207)通过表面等离子共振(MI：0107)(C)2010年欧洲生化学会联合会与NF-kappa-B p50(uniprotkb：P25799)和I-kappa-Bα(uniprotkb：O15111)物理相互作用。爱思唯尔出版，版权所有。
The factor inhibiting HIF-1 ( FIH-1) hydroxylates many ankyrin repeat-containing proteins including I kappa B alpha. It is widely speculated that hydroxylation of I kappa B alpha has functional consequences, but the effects of hydroxylation have not been demonstrated. We prepared hydroxylated I kappa B alpha and compared it to the unhydroxylated protein. Urea denaturation and amide H/D exchange experiments showed no change in the "foldedness" upon hydroxylation. Surface plasmon resonance measurements of binding to NF kappa B showed no difference in the NF kappa B binding kinetics or thermodynamics. Ubiquitin-independent proteasomal degradation experiments showed no difference in the half-life of the protein. Thus, it appears that hydroxylation of I kappa B alpha by FIH-1 is inconsequential, at least for the functions we could assay in vitro.Structured summary:MINT-8051494: NF-kappa-B p65 (uniprotkb:Q04207) physically interacts (MI: 0915) with NF-kappa-B p50 (uniprotkb:P25799) and I-kappa-B alpha (uniprotkb:O15111) by surface plasmon resonance (MI:0107) (c) 2010 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.