Biophysical approaches to solve the structures of the complex glycan shield of chloroviruses

Cristina De Castro, Garry A. Duncan, Domenico Garozzo, Antonio Molinaro, Luisa Sturiale, Michela Tonetti, James L. Van Etten

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

The capsid of Paramecium bursaria chlorella virus (PBCV-1) contains a heavily glycosylated major capsid protein, Vp54. The capsid protein contains four glycans, each N-linked to Asn. The glycan structures are unusual in many aspects: (1) they are attached by a β-glucose linkage, which is rare in nature; (2) they are highly branched and consist of 8–10 neutral monosaccharides; (3) all four glycoforms contain a dimethylated rhamnose as the capping residue of the main chain, a hyper-branched fucose residue and two rhamnose residues ''with opposite absolute configurations; (4) the four glycoforms differ by the nonstoichiometric presence of two monosaccharides, l-arabinose and d-mannose; (5) the N-glycans from all of the chloroviruses have a strictly conserved core structure; and (6) these glycans do not resemble any structures previously reported in the three domains of life. The structures of these N-glycoforms remained elusive for years because initial attempts to solve their structures used tools developed for eukaryotic-like systems, which we now know are not suitable for this noncanonical glycosylation pattern. This chapter summarizes the methods used to solve the chlorovirus complex glycan structures with the hope that these methodologies can be used by scientists facing similar problems.

Original languageEnglish (US)
Title of host publicationAdvances in Experimental Medicine and Biology
PublisherSpringer New York LLC
Pages237-257
Number of pages21
DOIs
StatePublished - 2018

Publication series

NameAdvances in Experimental Medicine and Biology
Volume1104
ISSN (Print)0065-2598
ISSN (Electronic)2214-8019

Keywords

  • GC-MS
  • Giant viruses
  • MALDI
  • N-glycosylation
  • NMR

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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