Thiogalactopyranosides are resistant to hydrolysis by α-galactosidases

Chembiochem. 2012 Jul 23;13(11):1673-9. doi: 10.1002/cbic.201200155. Epub 2012 Jun 27.

Abstract

Fluorescently tagged glycosides containing terminal α(1→3) and α(1→4)-linked thiogalactopyranosides have been prepared and tested for resistance to hydrolysis by α-galactosidases. Eight fluorescent glycosides containing either galactose or 5-thiogalactose as the terminal sugar were enzymatically synthesized using galactosyltransferases, with lactosyl glycosides as acceptors and UDP-galactose or UDP-5'-thiogalactose, respectively, as donors. The glycosides were incubated with human α-galactosidase A (CAZy family GH27, a retaining glycosidase), Bacteroides fragilis α-1,3-galactosidase (GH110, an inverting glycosidase), or homogenates of MCF-7 human breast cancer cells or NG108-15 rat glioma cells. Substrate hydrolysis was monitored by capillary electrophoresis with fluorescence detection. All compounds containing terminal O-galactose were readily degraded. Their 5-thiogalactose counterparts were resistant to hydrolysis by human α-galactosidase A and the enzymes present in the cell extracts. B. fragilis α-1,3-galactosidase hydrolyzed both thio- and O-galactoside substrates; however, the thiogalactosides were hydrolyzed at only 1-3 % of the rate of O-galactosides. The hydrolytic resistance of 5-thiogalactose was also confirmed by an in vivo study using cells in culture. The results suggest that 5-thiogalactosides may be useful tools for the study of anabolic pathways in cell extracts or in single cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacteroides fragilis / enzymology
  • Breast Neoplasms / metabolism*
  • Breast Neoplasms / pathology
  • Female
  • Glioma / metabolism*
  • Glioma / pathology
  • Humans
  • Hydrolysis
  • Molecular Structure
  • Rats
  • Thiogalactosides / chemical synthesis
  • Thiogalactosides / chemistry
  • Thiogalactosides / metabolism*
  • Tumor Cells, Cultured
  • alpha-Galactosidase / metabolism*

Substances

  • Thiogalactosides
  • alpha-Galactosidase