通过15N核磁共振确定锤头状核酶基序中两个位点上Mg2+、Zn2+和Cd2+的差异结合。

Differential binding of Mg2+, Zn2+, and Cd2+ at two sites in a hammerhead ribozyme motif, determined by 15N NMR.

作者信息

Wang Gang, Gaffney Barbara L, Jones Roger A

机构信息

Department of Chemistry and Chemical Biology, 610 Taylor Road, Rutgers, The State University of New Jersey, Piscataway, New Jersey 08854, USA.

出版信息

J Am Chem Soc. 2004 Jul 28;126(29):8908-9. doi: 10.1021/ja049804v.

DOI:10.1021/ja049804v

PMID:15264817

Abstract

A decamer duplex model of Domain II of the hammerhead ribozyme was synthesized with [8-13C-1,7,NH2-15N3]-guanosine at the known metal binding site, G10.1 and, for comparison, [2-13C-1,7,NH2-15N3]-guanosine at G16.2. The 15N NMR chemical shifts of the labeled N7s monitored during addition of Mg2+, Cd2+, and Zn2+ showed the same preference for binding at G10.1 over G16.2 for each metal. These results demonstrate that 15N labeling can be used to evaluate the binding of different metals, including Mg2+, to a given nitrogen, as well as to compare the binding potential of different sites.

摘要

在锤头状核酶结构域II的十聚体双链体模型中，在已知的金属结合位点G10.1处合成了带有[8-13C-1,7,NH2-15N3]-鸟苷的模型，作为比较，在G16.2处合成了[2-13C-1,7,NH2-15N3]-鸟苷。在添加Mg2+、Cd2+和Zn2+的过程中监测到的标记N7s的15N NMR化学位移表明，每种金属在G10.1处的结合偏好都高于G16.2处。这些结果表明，15N标记可用于评估不同金属（包括Mg2+）与给定氮的结合，以及比较不同位点的结合潜力。