FGF-FGFR signaling mediated through glycosaminoglycans in microtiter plate and cell-based microarray platforms

Biochemistry. 2013 Dec 17;52(50):9009-19. doi: 10.1021/bi401284r. Epub 2013 Dec 6.

Abstract

Fibroblast growth factor (FGF) signals cell growth through its interaction with a fibroblast growth factor receptor (FGFR) and a glycosaminoglycn (GAG) coreceptor. Here, we examine the signaling of five different FGFs (1, 2, 6, 8, and 8b) through FGFR3c. A small library of GAG and GAG-derivative coreceptors are screened to understand better the structure-activity relationship of these coreceptors on signaling. Initially, data were collected in a microtiter plate well-based cell proliferation assay. In an effort to reduce reagent requirements and improve assay throughput, a cell-based microarray platform was developed. In this cell-based microarray, FGFR3c-expressing cells were printed in alginate hydrogel droplets of ∼30 nL and incubated with FGF and GAG. Heparin was the most effective GAG coreceptor for all FGFs studied. Other GAGs, such as 2-O-desulfated heparin and chondroitin sulfate B, were also effective coreceptors. Signaling by FGF 8 and FGF 8b showed the widest tolerance for coreceptor structure. Finally, this on-chip cell-based microarray provides comparable data to a microtiter plate well-based assay, demonstrating that the coreceptor assay can be converted into a high-throughput assay.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Carbohydrate Conformation
  • Cell Proliferation
  • Cells, Cultured
  • Fibroblast Growth Factors / metabolism*
  • Glycosaminoglycans / chemistry
  • Glycosaminoglycans / metabolism*
  • Heparin / metabolism
  • Humans
  • Mice
  • Microarray Analysis*
  • Molecular Sequence Data
  • Receptors, Fibroblast Growth Factor / metabolism*
  • Recombinant Proteins / metabolism
  • Signal Transduction*

Substances

  • Glycosaminoglycans
  • Receptors, Fibroblast Growth Factor
  • Recombinant Proteins
  • Fibroblast Growth Factors
  • Heparin