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

Hyun Soo Lee; Jon S. Thorson

doi:10.1016/j.ab.2011.06.016

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

Hyun Soo Lee, Jon S. Thorson

Research output: Contribution to journal › Article › peer-review

22 Scopus citations

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.

Original language	English
Pages (from-to)	85-88
Number of pages	4
Journal	Analytical Biochemistry
Volume	418
Issue number	1
DOIs	https://doi.org/10.1016/j.ab.2011.06.016
State	Published - Nov 1 2011

Bibliographical note

Funding Information:
This work was supported by funding from the NIH ( AI52218 ) and the Laura and Edward Kremers Chair in Natural Products Chemistry (J.S.T.).

Keywords

Carbohydrate
Engineering
Enzyme
Evolution
Glycosyltransferase
Sugar nucleotide

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/j.ab.2011.06.016

Cite this

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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.",

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AU - Thorson, Jon S.

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N2 - 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.

AB - 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.

KW - Carbohydrate

KW - Engineering

KW - Enzyme

KW - Evolution

KW - Glycosyltransferase

KW - Sugar nucleotide

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JF - Analytical Biochemistry

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

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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