Using FRET to Study The Interaction Domain of TLR4 Binding to MD-2 in Living Cells

作者：Zhong Tian-Yu; Tang Jing; Chen Deng-Yu; Liu Ya-Wei; Wang Wei; Liu Jing-Hua^*; Jiang Yong

来源：Progress in Biochemistry and Biophysics, 2009, 36(11): 1451-1457.

DOI：10.3724/SP.J.1206.2009.00246

• MD-2
• TLR4
• LPS
• fluorescence resonance energy tramsfer(FRET)
• signal transduction

TLR4-MD-2 complex plays a key role in LPS recognition and its signal transduction. These steps are the vital elements of the host's defensive reaction. Studying the functional domain of TLR4 and MD-2 is very important to further understand the mechanism of LPS signal transduction. It was studied the interaction domain of TLR4 and MD-2 in living cells based on gene mutation, gene transfection and fluorescence resonance energy tramsfer(FRET) which is considered as one of the best methods used for intracellular protein-protein interaction study. CY-15P which was fused by CFP and YFP through 15 neutral amino acids was used as positive control, while co-expressed CFP and YFP proteins were used as negative control. The results showed that the ability of TLR4 binding to MD-2 decreased dramatically after the deletion of Glu(24) similar to Met(41) in N terminal of TLR4. Aggregation of TLR4 to LPS stimulation was observed, however, TLR4 without the Glu(24) similar to Met(41) mutation did not aggregate. All these results indicated that TLR4 Glu(24) similar to Met(41) might be the interaction domain of TLR4 binding to MD-2 and participate in the aggregation effect of TLR4 upon LPS stimulation.

• 出版日期2009-11
• 单位南方医科大学