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.
Highlights
The human HKU1 coronavirus (CoV) was first discovered in Hong Kong in 2004 and found to cause prevalent respiratory diseases (Woo et al, 2005)
Albumin bovine serum (BSA), trypsin, elastase, and chemical reagents of iodoacetamide (IAA), 1,4-dithiothreitol (DTT), acetic acid (HAc), ammonium bicarbonate (NH4HCO3), ammonia water (NH3·H2O), urea, and the zwitterionic hydrophilic interaction liquid chromatography (ZIC-Hydrophilic interaction liquid chromatography (HILIC)) materials were obtained from Sigma
The effect of different ACN contents (50–80%) on the O-glycopeptide enrichment on histidine-bonded silica (HBS) was investigated under the same pH condition
Summary
The human HKU1 coronavirus (CoV) was first discovered in Hong Kong in 2004 and found to cause prevalent respiratory diseases (Woo et al, 2005). Several β-CoVs, including mouse hepatitis virus, human CoV OC43, and bovine CoV (BCoV), use their NTDs to bind receptor protein (Peng et al, 2011; Peng et al, 2012). It was expected that HBS materials can be applied for the enrichment of O-glycopeptides from HKU1 S To achieve this goal, we first developed the enrichment method of O-glycopeptides based on HBS by optimizing different enrichment conditions with bovine fetuin as the model glycoprotein. We first developed the enrichment method of O-glycopeptides based on HBS by optimizing different enrichment conditions with bovine fetuin as the model glycoprotein This newly developed method was further validated by enriching O-glycopeptides from a mixture of bovine fetuin and albumin bovine serum digests, and commercial ZIC-HILIC materials were used for comparison. We believe that deciphering O-glycosylation will provide a significant complement to glycosylation for HKU1 S
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
