Bio-Inspired Liposomal Thrombomodulin Conjugate through Bio-Orthogonal Chemistry

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• 作者：Hailong Zhang ; Jacob Weingart ; Rui Jiang ; Jianhao Peng ; Qingyu Wu ; Xue-Long Sun
• 刊名：Bioconjugate Chemistry
• 出版年：2013
• 出版时间：April 17, 2013
• 年：2013
• 卷：24
• 期：4
• 页码：550-559
• 全文大小：480K
• 年卷期：v.24,no.4(April 17, 2013)
• ISSN：1520-4812

文摘

We report the synthesis of bioinspired liposomal thrombomodulin (TM) conjugates by chemoselective and site-specific liposomal conjugation of recombinant TM at C-terminus. TM is an endothelial cell membrane protein that acts as a major cofactor in the protein C anticoagulant pathway. To closely mimic membrane protein structural features of TM, we proposed membrane-mimetic re-expression of recombinant TM onto liposome. A recombinant TM containing the EGF-like 456 domains and an azidohomoalanine at C-terminus was expressed in E. coli. Conjugation of the recombinant TM onto liposome via Staudinger ligation and copper-free click chemistry were investigated as an optimal platform for exploring membrane protein TM鈥檚 activity, respectively. The bioinspired liposomal TM conjugates were confirmed with Western blotting and protein C activation activity. The recombinant TM-liposome conjugates showed a 2-fold higher k_cat/K_m value for protein C activation than that of the recombinant TM alone_, which indicated that the lipid membrane has a beneficiary effect on the recombinant TM鈥檚 activity. The reported liposomal protein conjugate approach provides a rational design strategy for both studying membrane protein TM鈥檚 functions and generating a membrane protein TM-based anticoagulant agent.