Synthesis of an End-to-End Protein–Glycopolymer Conjugate via Bio-Orthogonal Chemistry
Document Type
Article
Publication Date
1-19-2016
Publication Title
ACS Macro Letters
Abstract
We report the synthesis of an end-to-end protein–glycopolymer conjugate, namely, site-specific modification of recombinant thrombomodulin at the C-terminus with a chain-end-functionalized glycopolymer. Thrombomodulin (TM) is an endothelial membrane glycoprotein that acts as a major cofactor in the protein C anticoagulant pathway. To closely mimic the glycoprotein structural feature of native TM, we proposed a site-specific glyco-engineering of recombinant TM with a glycopolymer. Briefly, recombinant TM containing the epidermal growth factor (EGF)-like domains 4, 5, and 6 (rTM456) and a C-terminal azidohomoalanine was modified with a dibenzylcyclooctyne (DBCO) chain-end-functionalized glycopolymer via copper-free click chemistry to afford the end-to-end TM–glycopolymer conjugate. The TM glycoconjugation was confirmed with SDS-PAGE, Western blot, and protein C activation assay, respectively. The reported site-specific end-to-end protein glycopolymer conjugation approach facilitates uniform glycoconjugate formation via biocompatible chemistry and in high efficiency providing a rational strategy for generating an rTM-based anticoagulant agent.
Recommended Citation
Zhang, H.; Weingart, J.; Gruzdys, V.; Sun, X. Synthesis of an End-to-End Protein Glycopolymer Conjugate via Bio-Orthogonal Chemistry. Acs Macro Letters 2016, 5, 78-82.
DOI
10.1021/acsmacrolett.5b00805
Volume
5
Issue
1
Comments
This work was supported in part by grants from the American Heart Association (14GRNT20290002, X.-L. Sun), NIH (1R01HL102604-04, X.-L. Sun), National Science Foundation (CHE-1126384, X.-L. Sun), and Cleveland State University Center for Gene Regulation in Health and Disease Fund.