Efficient Chemical Synthesis and Folding of Mirror-Image Tropomyosin Receptor Kinase A Using the Strategy of Removable Glycosylation Modification

Abstract

The strategy of removable glycosylation modification was used to overcome the low-efficiency problem encountered in the chemical synthesis of the mirror-image D-version of the immunoglobulin (Ig)-like domain of tropomyosin receptor kinase A ( ^DlgC ^TrkA), a protein molecule needed for mirror-image screening of D-peptide ligands targeting this cell membrane receptor. It was found that O-linked-β- N-acetyl- D-glucosamine (O-GlcNAc) modification at ^DSer ³¹², or ^DSer ³²⁰ can significantly improve the efficiency of ^DlgC ^TrkA synthesis and folding, while O-GlcNAc modification at ^DSer ³³⁰ showed barely any improvement. This study provides a new example demonstrating the power of the removable glycosylation modification strategy in the chemical synthesis and folding of difficult-to-obtain proteins. It also presents evidence that removable glycosylation modification at different sites would significantly affect the efficiency of protein folding promoted by this strategy.

Keywords

Chemical protein synthesis / Removable glycosylation modification / Synthetic methods / Protein folding / D-protein

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2024 SIOC, CAS, Shanghai, & WILEY-VCH GmbH