Abstract
Oligosaccharyltransferase (OT) catalyzes the signature reaction of the asparagine-linked glycosylation pathway, namely, the transfer of preformed glycans from the lipid-linked oligosaccharide Glc3Man9GlcNAc2-P-P-Dolichol (G3M9Gn2-LLO) to appropriate asparaginyl residues on acceptor polypeptides. We have identified a reaction, possibly catalyzed by OT, that results in the hydrolysis or “transfer to water” of host LLOs in response to viral infection with release of a free G3M9Gn2 glycan. The loss of LLO ostensibly hinders N-glycosylation of viral polypeptides. This response is achieved by a novel stress-activated signaling pathway in which free mannose-6-phosphate (M6P) acts as a second-messenger. Here, we describe methods with permeabilized mammalian cells for activation of the M6P-regulated LLO hydrolysis, or transfer of glycan to water, in vitro.
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References
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Acknowledgments
This work was supported by generous funding from the National Institutes of General Medical Sciences (NIH, grant GM38545) and the Robert Welch Foundation (grant I-1168).
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Gao, N., Lehrman, M.A. (2013). Mannose-6-Phosphate: A Regulator of LLO Destruction. In: Brockhausen, I. (eds) Glycosyltransferases. Methods in Molecular Biology, vol 1022. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-465-4_20
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DOI: https://doi.org/10.1007/978-1-62703-465-4_20
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-464-7
Online ISBN: 978-1-62703-465-4
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