Washington Chromatography Discussion Group (WCDG) meets monthly from September to May. Meetings are free and include a light dinner & social hour followed by a seminar.
September 16, 2020
Speaker: Asif Shajahan, University of Georgia
Title: How advanced mass spectrometry can guide the therapeutic interventions against SARS-CoV-2
Webinar with breakout networking sessions
Abstract: The development of new therapeutic strategies against COVID-19 pandemic emergence caused by SARS-CoV-2 requires understanding of mode of viral attachment, entry, and replication. We are focused on the glycobiology of SARS-CoV-2 and on how it can help both in development of vaccine and therapeutics. The heavily glycosylated coronavirus surface spike (S) protein facilitates viral attachment, entry and membrane fusion and also plays a critical role in the elicitation of host immune response. The SARS-CoV-2 spike protein is comprised of two protein subunits (S1 and S2) and together possess 22 potential N-glycosylation sites. The S proteins of SARS-CoV-1 and SARS-CoV-2 primarily binds to human angiotensin converting enzyme 2 (hACE2) for the host cell entry. Even though hACE2 has been known for two decades and has been recognized as the entry point of several human coronaviruses, comprehensive glycosylation data of hACE2 is currently not available. Herein, we describe the quantitative N- and O- glycosylation mapping on S protein subunits S1 and S2, and also hACE2 expressed in human cells by both glycoproteomics and glycomics via high resolution LC-MSn. We have characterized the N-glycosylation profile on S protein and interestingly, observed an unexpected O-glycosylation modification on the receptor binding domain (RBD) of spike protein subunit S1. This is the first report of experimental data for both the site of O-glycosylation and identity of the O-glycans attached on the subunit S1 of SARS-CoV-2. We observed almost complete glycan occupancy at all seven possible N-glycosylation sites on hACE2 along with detection of three novel O-glycosylation sites. We have characterized the terminal sialic acid linkages, the presence of bisecting GlcNAc, and the pattern of N-glycan fucosylation to understand the detailed structure of glycan epitopes on hACE2 involved in viral binding. Understanding the site-specific glycosylation and terminal glycan epitopes on viral spike protein and its receptor hACE2 receptor can aid in the development of novel therapy.
Bio: Dr. Shajahan has about 10 years of experience in the structural characterization of glycoproteins using state-of-the-art high-resolution mass spectrometry (MS). He completed his Ph.D. in 2014 where he worked in the interface of chemistry and biology by developing glycoconjugates for the glycoengineering of brain glycans (across BBB) in mice models. He joined CCRC, University of Georgia, in 2015 and have been doing characterization of glycoproteins by both glycomics and glycoproteomics. He has contributed to over 80 projects on the structural characterization of mammalian, plant, algal, insect and bacterial origin glycoconjugates from both academic and industrial researchers. He is also performing research for the development of novel methods which enables rapid but comprehensive glycosylation profiling. He is training and mentoring undergraduate students and other junior postdoctoral trainees at CCRC and also involved as an instructor for the annual hand-on training courses held at CCRC on MS based glycomics and glycoproteomics. He is currently working as an Assistant Research Scientist faculty at CCRC.
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