Isolation of bovine corneal keratan sulfate and its growth factor and morphogen binding

FEBS J. 2013 May;280(10):2285-93. doi: 10.1111/febs.12165. Epub 2013 Feb 28.

Abstract

Keratan sulfate (KS) is an important glycosaminoglycan that is found in cartilage, reproductive tissues, and neural tissues. Corneal KS glycosaminoglycan is found N-linked to lumican, keratocan and mimecan proteoglycans, and has been widely studied by investigators interested in corneal development and diseases. Recently, the availability of corneal KS has become severely limited, owing to restrictions on the shipment of bovine central nervous system byproducts across international borders in an effort to prevent additional cases of mad cow disease. We report a simple method for the purification of multi-milligram quantities of bovine corneal KS, and characterize its structural properties. We also examined its protein-binding properties, and discovered that corneal KS bound with high affinity to fibroblast growth factor-2 and sonic hedgehog, a growth factor and a morphogen involved in corneal development and healing.

Publication types

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

MeSH terms

  • Animals
  • Cattle
  • Chromatography, High Pressure Liquid / methods
  • Cornea / chemistry*
  • Fibroblast Growth Factor 1 / chemistry
  • Fibroblast Growth Factor 2 / chemistry
  • Hedgehog Proteins / chemistry*
  • Keratan Sulfate / chemistry
  • Keratan Sulfate / isolation & purification*
  • Magnetic Resonance Spectroscopy
  • Molecular Structure
  • Molecular Weight
  • Protein Binding
  • Protein Interaction Mapping / methods*

Substances

  • Hedgehog Proteins
  • Fibroblast Growth Factor 2
  • Fibroblast Growth Factor 1
  • Keratan Sulfate