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

Matthew Attreed, Hannes E. Buelow

Research output: Contribution to journalArticle

2 Citations (Scopus)

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 (Clifton, N.J.)
Volume1229
DOIs
StatePublished - 2015
Externally publishedYes

Fingerprint

Heparitin Sulfate
Cations
Single-Chain Antibodies
Glycosaminoglycans
Protein Binding
Extracellular Matrix
Epitopes
Antibodies

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

A transgenic approach to live imaging of heparan sulfate modifi cation patterns. / Attreed, Matthew; Buelow, Hannes E.

In: Methods in molecular biology (Clifton, N.J.), Vol. 1229, 2015, p. 253-268.

Research output: Contribution to journalArticle

@article{7bc635f8e4eb4d13aa0437d0d88ebfb8,
title = "A transgenic approach to live imaging of heparan sulfate modifi cation patterns",
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.",
keywords = "Caenorhabditis elegans, Heparan sulfate, Live imaging, Nongenetically encoded molecules, Single chain variable fragment (scFv) antibody",
author = "Matthew Attreed and Buelow, {Hannes E.}",
year = "2015",
doi = "10.1007/978-1-4939-1714-3_22",
language = "English (US)",
volume = "1229",
pages = "253--268",
journal = "Methods in Molecular Biology",
issn = "1064-3745",
publisher = "Humana Press",

}

TY - JOUR

T1 - A transgenic approach to live imaging of heparan sulfate modifi cation patterns

AU - Attreed, Matthew

AU - Buelow, Hannes E.

PY - 2015

Y1 - 2015

N2 - 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.

AB - 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.

KW - Caenorhabditis elegans

KW - Heparan sulfate

KW - Live imaging

KW - Nongenetically encoded molecules

KW - Single chain variable fragment (scFv) antibody

UR - http://www.scopus.com/inward/record.url?scp=84921714492&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84921714492&partnerID=8YFLogxK

U2 - 10.1007/978-1-4939-1714-3_22

DO - 10.1007/978-1-4939-1714-3_22

M3 - Article

C2 - 25325959

AN - SCOPUS:84921714492

VL - 1229

SP - 253

EP - 268

JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

SN - 1064-3745

ER -