Deciphering the O-Glycosylation of HKU1 Spike Protein With the Dual-Functional Hydrophilic Interaction Chromatography Materials

Yun Cui; Xuefang Dong; Xiaofei Zhang; Cheng Chen; Dongmei Fu; Xiuling Li; Xinmiao Liang

doi:10.3389/fchem.2021.707235

Deciphering the O-Glycosylation of HKU1 Spike Protein With the Dual-Functional Hydrophilic Interaction Chromatography Materials

Front Chem. 2021 Aug 13:9:707235. doi: 10.3389/fchem.2021.707235. eCollection 2021.

Authors

Yun Cui¹, Xuefang Dong², Xiaofei Zhang², Cheng Chen², Dongmei Fu¹, Xiuling Li², Xinmiao Liang²

Affiliations

¹ School of Biological Engineering, Dalian Polytechnic University, Dalian, China.
² Key Lab of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.

Abstract

HKU1 is a human beta coronavirus and infects host cells via highly glycosylated spike protein (S). The N-glycosylation of HKU1 S has been reported. However, little is known about its O-glycosylation, which hinders the in-depth understanding of its biological functions. Herein, a comprehensive study of O-glycosylation of HKU1 S was carried out based on dual-functional histidine-bonded silica (HBS) materials. The enrichment method for O-glycopeptides with HBS was developed and validated using standard proteins. The application of the developed method to the HKU1 S1 subunit resulted in 46 novel O-glycosylation sites, among which 55.6% were predicted to be exposed on the outer protein surface. Moreover, the O-linked glycans and their abundance on each HKU1 S1 site were analyzed. The obtained O-glycosylation dataset will provide valuable insights into the structure of HKU1 S.

Keywords: HKU1; O-glycosylation abundance; O-glycosylation sites; enrichment; spike glycoprotein.