Synthesis of an End-to-End Protein–Glycopolymer Conjugate via Bio-Orthogonal Chemistry

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• 作者：Hailong Zhang ; Jacob Weingart ; Valentinas Gruzdys ; Xue-Long Sun
• 刊名：ACS Macro Letters
• 出版年：2016
• 出版时间：January 19, 2016
• 年：2016
• 卷：5
• 期：1
• 页码：73-77
• 全文大小：353K
• ISSN：2161-1653

文摘

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.