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	Analysis of beryllium to biomolecule binding using a metal specific fluorescent probe and competitive assay
	Zheng, Yan ; Lin, Lin ; Hang, Wei ; Hang W(杭纬) ; Yan, Xiaomei ; Yan XM(颜晓梅) ; Marrone, Babetta L.（Los Alamos Natl Lab, Biosci Div）
	2011-04-16
关键词	Beryllium Dissociation constant Metal Fluorescence Nucleotides
英文摘要	Studying metal-biomolecule interactions is critical to the elucidation of the molecular basis of the biological functions and toxicity of metals. In the present study, a competitive fluorimetric approach has been developed to measure the apparent affinity of biomolecules for Be(2+) by using a Be(2+)-specific fluorigenic probe (10-hydroxybenzo[h]quinoline-7-sulfonate, HBQS). Under physiological conditions, HBQS coordinates with Be(2+) in a molar ratio of 1:1 and results in a fluorescence shift from 580 nm for HBQS to 480 nm for the Be-HBQS complex associated with significant fluorescence enhancement. When a beryllium ligand is present in the mixture of Be(2+) and HBQS, the competition of ligand against HBQS for beryllium ion binding results in dissociation and thus a fluorescence decrease of the Be-HBQS complex. By titrating ligand and monitoring the dose-dependent decrease of Be-HBQS complex fluorescence at 480 nm, the apparent affinity between ligand and Be(2+) can be derived. Applying this simple approach, the apparent affinities of various nucleotides and the iron-storage protein ferritin for beryllium ion have been determined. In particular, the apparent dissociation constant of Be(2+) and adenosine 5'-triphosphate (ATP) was also validated by an electrospray ionization mass spectrometric (ESI-MS) method. The general applicability of the proposed competition assay was further demonstrated using FluoZin-1, a zinc fluorescent indicator, in a binding study for Zn(2+) and bovine serum albumin. This newly developed competitive fluorimetric assay provides a sensitive, simple, and generic approach for affinity estimation of metal and biomolecule binding. (C) 2011 Elsevier B.V. All rights reserved.; program for New Century Excellent Talents in University[NCET-07-0729]; Los Alamos National Laboratory
语种	英语
出版者	ELSEVIER SCIENCE BV
内容类型	期刊论文
源URL	[http://dx.doi.org/doi:10.1016/j.talanta.2011.04.037]
专题	化学化工－已发表论文
推荐引用方式 GB/T 7714	Zheng, Yan,Lin, Lin,Hang, Wei,et al. Analysis of beryllium to biomolecule binding using a metal specific fluorescent probe and competitive assay[J],2011.
APA	Zheng, Yan.,Lin, Lin.,Hang, Wei.,杭纬.,Yan, Xiaomei.,...&Marrone, Babetta L..(2011).Analysis of beryllium to biomolecule binding using a metal specific fluorescent probe and competitive assay..
MLA	Zheng, Yan,et al."Analysis of beryllium to biomolecule binding using a metal specific fluorescent probe and competitive assay".(2011).

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