Bio-Inspired Liposomal Thrombomodulin Conjugate through Bio-Orthogonal Chemistry
Document Type
Article
Publication Date
4-2013
Publication Title
Bioconjugate Chemistry
Abstract
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 higher kcat/Km 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’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.
Recommended Citation
Zhang, H.; Weingart, J.; Jiang, R.; Peng, J.; Wu, Q.; Sun, X. Bio-Inspired Liposomal Thrombomodulin Conjugate through Bio-Orthogonal Chemistry. Bioconjugate Chem. 2013, 24, 550-559.
DOI
10.1021/bc300399f
Volume
24
Issue
4
Comments
This work was supported by grants from the NIH
(1R01HL102604-04, X.-L. Sun), the Ohio Research Scholar Program (X.-L. Sun and Q. Wu), National Science Foundation MRI Grant (CHE-1126384, X.-L. Sun), and Cleveland State University Faculty Research Development Grant.