Glycosylation profiling of monkeypox virus structural proteins with poly Ser-Arg materials

Area of Science:

• Virology
• Glycobiology
• Materials Science

Background:

• Viral protein glycosylation is crucial for host cell invasion.
• The glycosylation patterns of monkeypox virus (MPXV) structural proteins remain largely uncharacterized.

Purpose of the Study:

• To investigate the role of glycosylation in MPXV infection.
• To develop and validate novel materials for comprehensive glycopeptide enrichment.
• To analyze the N-glycosylation and O-glycosylation of key MPXV structural proteins.

Main Methods:

• Synthesis of poly Ser-Arg (poly SR) materials via surface-initiated reversible addition-fragmentation chain transfer (SI-RAFT) polymerization.
• Evaluation of poly SR materials using standard proteins and complex biological lysates (bovine, mouse, human).
• Application of poly SR materials for comprehensive N- and O-glycopeptide analysis of MPXV structural proteins (A29, A35, B6R, H3L) and glycan microarray analysis of protein A29 binding.

Main Results:

• Poly SR materials demonstrated excellent enrichment performance for both N- and O-glycopeptides.
• MPXV structural proteins A29, A35, B6R, and H3L were found to be highly sialylated.
• MPXV protein A29 exhibited strong specific binding to heparan sulfate and chondroitin sulfate.

Conclusions:

• The developed poly SR materials are effective for comprehensive glycopeptide analysis.
• High sialylation of MPXV structural proteins suggests a significant role in viral infection.
• Specific binding of MPXV protein A29 to host glycosaminoglycans provides insights into viral entry mechanisms.
• Findings lay the groundwork for developing MPXV vaccines and antiviral drugs.