Fluorescent SNAP-Tag Galectin Fusion Proteins as Novel Tools in Glycobiology

Christiane E. Kupper, Sophia Boecker, Hulong Liu, Carina Adamzyk, Julia van de Kamp, Tobias Recker, Bernd Lethaus, Willi Jahnen-Dechent, Sabine Neuss, Gerhard Mueller-Newen, Lothar Elling*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

15 Citations (Web of Science)

Abstract

Galectins,??-galactoside binding proteins, function in several physiological and pathological processes. The further evaluation of these processes as well as possible applications of galectins in diagnosis and therapy has raised high scientific interest. Therefore, easy and reliable test systems are necessary. Here we present the simple and cost-efficient production of recombinant human galectins as fusion proteins with SNAP-tag and fluorescent proteins. These constructs show binding specificities and oligomerisation properties generally comparable to recombinant galectins. Their direct fluorescence signal was utilised by ELISA-type assay and flow cytometry analysis with human and ovine mesenchymal stem cells (MSC). Flow cytometry demonstrated glycan mediated binding of His6-SNAP-YFP-Gal- 3 to both MSC types, which was specifically inhibited by lactose. Moreover, directed immobilisation by SNAP-tag technology onto benzylguanine- activated sepharose was utilised to prepare galectin affinity columns for glycoprotein analysis and purification. The SNAPtag directed coupling yielded up to three-fold higher binding capacities for the glycoprotein standard asialofetuin compared to nondirected coupled galectin suggesting improved functionality following directed coupling.
Original languageEnglish
Pages (from-to)5457-5467
JournalCurrent Pharmaceutical Design
Volume19
Issue number30
DOIs
Publication statusPublished - Sep 2013

Keywords

  • Galectin-1
  • galectin-3
  • SNAP-tag
  • fusion protein
  • fluorescent protein
  • mesenchymal stem cells

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