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中国生物工程杂志

China Biotechnology
China Biotechnology  2011, Vol. 31 Issue (10): 12-17    DOI:
    
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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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 wordsIRAK2      TLR      Inflammatory response     
Received: 20 May 2011      Published: 25 October 2011
ZTFLH:  Q819  
Cite this article:

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.

URL:

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


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