Imaging Glycosaminoglycan Modification Patterns In Vivo

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Glycosaminoglycans (GAGs) such as heparan sulfates (HS) or chondroitin sulfates (CS) are long unbranched polymers of a disaccharide comprised of hexuronic acid and hexosamine. Attached to a protein backbone via a characteristic tetrasaccharide, the GAG chains are non-uniformly modified by sulfations, epimerizations, and deacetylations. The resultant glycan chains contain highly modified domains, separated by sections of sparse or no modifications. These GAG domains are central to the role of glycans in binding to proteins and mediating protein–protein interactions. Since HS and CS domains are not genetically encoded, they cannot be visualized and studied with conventional methods in vivo. We describe a transgenic approach using single chain variable fragment (scFv) antibodies that bind HS or CS. By transgenically expressing fluorescently tagged scFv antibodies, we can directly visualize both HS and CS domains in live Caenorhabditis elegans revealing unprecedented cellular specificity and evolutionary conservation (Attreed et al., Nat Methods 9(5): 477–479, 2012; Attreed et al., Glycobiology 26(8): 862–870, 2016) (unpublished). The approach allows concomitant co-labeling of multiple GAG domains, the study of GAG dynamics, and could lend itself to a genetic analysis of GAG domain biosynthesis or function.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages539-557
Number of pages19
DOIs
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2303
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Caenorhabditis
  • Chondroitin sulfate
  • Dermatan sulfate
  • Glycosaminoglycans
  • Heparan sulfate
  • Live imaging
  • Non-genetically encoded molecules
  • Single chain variable fragment (scFv) antibody

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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