A transgenic approach to live imaging of heparan sulfate modifi cation patterns

Matthew Attreed, Hannes E. Bülow

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Heparan sulfate (HS) glycosaminoglycan chains contain highly modifi ed HS domains that are separated by sections of sparse or no modifi cation. HS domains are central to the role of HS in protein binding and mediating protein–protein interactions in the extracellular matrix. Since HS domains are not genetically encoded, they are impossible to visualize and study with conventional methods in vivo. Here we describe a transgenic approach using previously described single chain variable fragment (scFv) antibodies that bind HS in vitro and on tissue sections with different specifi cities. By engineering a secretion signal and a fl uorescent protein to the scFvs and transgenically expressing these fl uorescently tagged antibodies in aenorhabditis elegans, we are able to directly visualize specifi c HS domains in live animals (Attreed et al. Nat Methods 9(5):477–479, 2012). The approach allows concomitant colabeling of multiple epitopes, the study of HS dynamics and, could lend itself to a genetic analysis of HS domain biosynthesis or to visualize other nongenetically encoded or posttranslational modifi cations.

Original languageEnglish (US)
Pages (from-to)253-268
Number of pages16
JournalMethods in Molecular Biology
Volume1229
DOIs
StatePublished - Jan 1 2015
Externally publishedYes

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Keywords

  • Caenorhabditis elegans
  • Heparan sulfate
  • Live imaging
  • Nongenetically encoded molecules
  • Single chain variable fragment (scFv) antibody

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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