Bio-inspired liposomal thrombomodulin conjugate through bio-orthogonal chemistry.
Bio-inspired liposomal thrombomodulin conjugate through bio-orthogonal chemistry.
复制标题
通过生物正交化学仿生脂质体血栓调节蛋白缀合物。
DOI：
10.1021/bc300399f
复制
发表时间：
2013
影响因子：
4.7
通讯作者：
Sun,Xue-Long
中科院分区：
文献类型：
--
作者：
Zhang,Hailong;Weingart,Jacob;Jiang,Rui;Peng,Jianhao;Wu,Qingyu;Sun,Xue-Long
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’s 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 higherkcat/Kmvalue 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’s activity. The reported liposomal protein conjugate approach provides a rational design strategy for both studying membrane protein TM’s functions and generating a membrane protein TM-based anticoagulant agent.
