Ninjurin1 regulates lipopolysaccharide-induced inflammation through direct binding

Shin,Min  Wook; Bae,Sung-Jin; Wee,Hee-Jun; Lee,Hyo-Jong; Ahn,Bum  Ju; Le,Hoang; Lee,Eun  Ji; Kim,Ran  Hee; Lee,Hye  Shin; Seo,Ji  Hae; Park,Ji-Hyeon; Kim,Kyu-Won

doi:10.3892/ijo.2015.3296

Ninjurin1 regulates lipopolysaccharide-induced inflammation through direct binding

Authors:
- Min Wook Shin
- Sung-Jin Bae
- Hee-Jun Wee
- Hyo-Jong Lee
- Bum Ju Ahn
- Hoang Le
- Eun Ji Lee
- Ran Hee Kim
- Hye Shin Lee
- Ji Hae Seo
- Ji-Hyeon Park
- Kyu-Won Kim
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Affiliations: SNU-Harvard NeuroVascular Protection Research Center, College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul 151-742, Republic of Korea, College of Pharmacy, Inje University, Gimhae, Gyugnam 621-749, Republic of Korea
Published online on: December 15, 2015 https://doi.org/10.3892/ijo.2015.3296
Pages: 821-828

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Abstract

Ninjurin1 is a transmembrane protein involved in macrophage migration and adhesion during inflammation. It was recently reported that repression of Ninjurin1 attenuated the lipopolysaccharide (LPS)-induced inflammatory response in macrophages; however, the precise mechanism by which Ninjurin1 modulates LPS-induced inflammation remains poorly understood. In the present study, we found that the interaction between Ninjurin1 and LPS contributed to the LPS-induced inflammatory response. Notably, pull-down assays using lysates from HEK293T cells transfected with human or mouse Ninjurin1 and biotinylated LPS (LPS-biotin) showed that LPS directly bound Ninjurin1. Subsequently, LPS binding assays with various truncated forms of Ninjurin1 protein revealed that amino acids (aa) 81-100 of Ninjurin1 were required for LPS binding. In addition, knockdown experiments using Ninj1 siRNA resulted in decreased nitric oxide (NO) and tumor necrosis factor-α (TNFα) secretion upon LPS treatment in Raw264.7 cells. Collectively, our results suggest that Ninjurin1 regulates the LPS-induced inflammatory response through its direct binding to LPS, thus, identifying Ninjurin1 as a putative target for the treatment of inflammatory diseases, such as sepsis and inflammation-associated carcinogenesis.

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