开发一种适用于高通量格式的通用糖基转移酶测定法。
Development of a universal glycosyltransferase assay amenable to high-throughput formats.
机构信息
Pharmaceutical Sciences Division, School of Pharmacy, Wisconsin Center for Natural Products Research, University of Wisconsin-Madison, Madison, WI 53705, USA.
出版信息
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.
摘要
本文描述了一种通用的 1-Zn(II)核苷二磷酸（NDP）传感器测定法的开发，用于快速评估糖基转移酶（GT）的活性。1-Zn(II) NDP 传感器测定法具有亚微摩尔的灵敏度，与纯化酶和粗细胞提取物兼容，并对 NDP 具有极高的选择性，超过相应的 NDP-糖。因此，预计 1-Zn(II) NDP 传感器测定法将在 GT 工程和表征方面具有广泛的适用性。
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