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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2011, Vol. 31 Issue (10): 12-17    
研究报告     
白细胞介素-1受体相关激酶-2的共价修饰与激酶活性的研究
尹卫国1,2, 高媛1, 梁瑜1, 李映菊1, 肖建华1
1. 南华大学病原生物学研究所 衡阳 421001;
2. 克利夫兰医学中心免疫系 俄亥俄州克利夫兰市 44195
Study on the Modifications and Kinase Activity of IRAK2
YIN Wei-guo1,2, GAO Yuan1, LIANG Yu1, LI Ying-ju1, XIAO Jian-hua1
1. Pathogenic Biology Institute, University of South China, Hengyang 421001, China;
2. Department of Immunology, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio 44195, U.S.A
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摘要:

白细胞介素-1受体相关激酶-2(IRAK2)是调节IL1和Toll样受体信号通路的一个关键性分子,目前对于共价修饰如何调节IRAK2的活性还所知甚少。当与IRAK1共转染时,IRAK2能被共价修饰,在SDS-PAGE分离中呈现出迁移率变慢的多条电泳带。在小鼠骨髓干细胞分化的巨噬细胞(BMMs)中,内源表达的IRAK2在TLR配体刺激下也呈现出类似的共价修饰。而且IRAK2的共价修饰具有磷酸酯酶敏感性,提示大部分为磷酸化修饰。通过体外磷酸激酶活性分析,发现巨噬细胞中表达的IRAK2能在LPS诱导下被激活,成为一个具有激酶活性的调节蛋白。进一步研究发现激酶灭活的IRAK2突变体不能重建IRAK2基因敲除巨噬细胞的功能。通过Western杂交和定量PCR分析,发现IRAK2的激酶活性是介导LPS诱导的信号通路和炎症因子表达所必须的。因此,在LPS诱导下,IRAK2可能被IRAK1进行磷酸化修饰而活化,从而介导下游的信号转导通路、诱导炎症因子的表达。

关键词: 白细胞介素-1受体相关激酶-2Toll样受体炎症反应    
Abstract:

IRAK2 plays a critical role in IL1 and TLR-mediated signaling, however, the mechanisms by which IRAK2 regulates IL1/TLR signaling remain unclear. It is found that IRAK2 became modified by co-transfected IRAK1 in 293 cells. The modified IRAK2 were recognized by anti-IRAK2 Western blotting as slower mobility shift bands on SDS-PAGE. In bone-marrow derived macrophages, endogenous IRAK2 was also modified upon LPS induction. Moreover, the modified IRAK2 bands collapsed upon pretreatment by calf intestinal phosphatase, indicating that IRAK2 modification is mainly attributed to phosphorylation. By in vitro kinase assay, immunoprecipitated IRAK2 was demonstrated to be able to phosphorylate MBP in vitro, suggesting that LPS signaling induces the kinase activity of IRAK2. Furthermore, a kinase-inactive mutant of IRAK2 is made, which is unable to reconstitute the LPS-induced signaling and inflammatory gene expression in IRAK2-deficient macrophages. Taken together, it demonstrates that the kinase activity of IRAK2 is critical for LPS-induced downstream signaling and proinflammatory gene expression, in addition, phosphorylation of IRAK2, likely regulated by IRAK1 might have a key role in activating IRAK2 kinase.

Key words: IRAK2    TLR    Inflammatory response
收稿日期: 2011-05-20 出版日期: 2011-10-25
ZTFLH:  Q819  
基金资助:

国家自然科学基金(30972576);湖南省科技厅科研基金(2010SK3038)资助项目

通讯作者: 肖建华     E-mail: jhxiao223@163.com
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引用本文:

尹卫国, 高媛, 梁瑜, 李映菊, 肖建华. 白细胞介素-1受体相关激酶-2的共价修饰与激酶活性的研究[J]. 中国生物工程杂志, 2011, 31(10): 12-17.

YIN Wei-guo, GAO Yuan, LIANG Yu, LI Ying-ju, XIAO Jian-hua. Study on the Modifications and Kinase Activity of IRAK2. China Biotechnology, 2011, 31(10): 12-17.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/        https://manu60.magtech.com.cn/biotech/CN/Y2011/V31/I10/12


[1] Wang C, Deng L, Hong M, et al. TAK-1 is a ubiquitin-dependent kinase of MKK and IKK. Nature, 2001, 412(6844):346-351.

[2] Yi A K, Peckham D W, Ashman R F, et al.CpG DNA rescues B cells from apoptosis by activating NFkappaB and preventing mitochondrial membrane potential disruption via a chloroquine-sensitive pathway. Int Immunol, 1999, 11(12):2015-2024.

[3] Deng L, Wang C, Spencer E, et al. Activation of the IkappaB kinase complex by TRAF6 requires a dimeric ubiquitin-conjugating enzyme complex and a unique polyubiquitin chain.Cell, 2000, 103(2):351-361.

[4] Ninomiya-Tsuji J, Kishimoto K, Hiyama A, et al.The kinase TAK1 can activate the NIK-I kappaB as well as the MAP kinase cascade in the IL-1 signalling pathway. Nature, 1999, 398(6724):252-256.

[5] Kim T W, Staschke K, Bulek K, et al. A critical role for IRAK4 kinase activity in Toll-like receptor-mediated innate immunity. J Exp Med, 2007, 204(5):1025-1036.

[6] Dunne A, Carpenter S, Brikos C, et al. IRAK1 and IRAK4 promote phosphorylation, ubiquitination, and degradation of MyD88 adaptor-like (Mal). J Biol Chem, 2010, 285(24):18276-18282.

[7] Wesche H, Gao X, Li X, et al. IRAK-M is a novel member of the Pelle/interleukin-1 receptor-associated kinase (IRAK) family. J Biol Chem, 1999, 274(27):19403-19410.

[8] Muzio M, Ni J, Feng P, et al. IRAK (Pelle) family member IRAK-2 and MyD88 as proximal mediators of IL-1 signaling. Science, 1997, 278(5343):1612-1615.

[9] Kawagoe T, Sato S, Matsushita K, et al. Sequential control of Toll-like receptor-dependent responses by IRAK1 and IRAK2. Nat Immunol, 2008, 9(6):684-691.

[10] Wan Y, Xiao H, Affolter J, et al.Interleukin-1 receptor-associated kinase 2 is critical for lipopolysaccharide-mediated post-transcriptional control. J Biol Chem, 2009, 284(16):10367-10375.

[11] Huang Y, Li T, Sane D C, et al. IRAK1 serves as a novel regulator essential for lipopolysaccharide-induced interleukin-10 gene expression. J Biol Chem, 2004, 279(49):51697-51703.

[12] Kollewe C, Mackensen A C.Sequential autophosphorylation steps in the interleukin-1 receptor-associated kinase-1 regulate its availability as an adapter in interleukin-1 signaling. J Biol Chem, 2004, 279(7):5227-5236.

[13] Caput D, Beutler B, Hartog K, et al.Identification of a common nucleotide sequence in the 3'-untranslated region of mRNA molecules specifying inflammatory mediators. Proc Natl Acad Sci U S A, 1986, 83(6):1670-1674.

[14] Shaw G, Kamen R. A conserved AU sequence from the 3' untranslated region of GM-CSF mRNA mediates selective mRNA degradation. Cell, 1986, 46(5):659-667.

[15] Hartupee J, Li X, Hamilton T. Interleukin 1α-induced NF-κB activation and chemokine mRNA stabilization diverge at IRAK1. J Biol Chem, 2008, 283(23):15689-15693.

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