Development of a universal glycosyltransferase assay amenable to high-throughput formats

Anal Biochem. 2011 Nov 1;418(1):85-8. doi: 10.1016/j.ab.2011.06.016. Epub 2011 Jul 8.

Abstract

The development of a general 1-Zn(II) nucleoside diphosphate (NDP) sensor assay for rapid evaluation of glycosyltransferase (GT) activity is described. The 1-Zn(II) NDP sensor assay offers submicromolar sensitivity, compatibility with both purified enzymes and crude cell extracts, and exquisite selectivity for NDPs over the corresponding NDP-sugars. Thus, the 1-Zn(II) NDP sensor assay is anticipated to offer broad applicability in the context of GT engineering and characterization.

Publication types

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

MeSH terms

  • Biological Assay
  • Biosensing Techniques
  • Coordination Complexes / chemistry*
  • Dinucleoside Phosphates / chemistry*
  • Glycosyltransferases / chemistry*
  • Glycosyltransferases / metabolism
  • High-Throughput Screening Assays
  • Kinetics
  • Zinc*

Substances

  • Coordination Complexes
  • Dinucleoside Phosphates
  • Glycosyltransferases
  • Zinc