Fluorescence spectroscopic study of serum albumin-bromadiolone interaction: fluorimetric determination of bromadiolone
Fluorescence spectroscopic study of serum albumin-bromadiolone interaction: fluorimetric determination of bromadiolone
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DOI：
10.1016/j.jpba.2005.01.023
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发表时间：
2005-07-01
影响因子：
3.4
通讯作者：
Mishra, AK
中科院分区：
文献类型：
--
作者：
Deepa, S;Mishra, AK
溴敌隆（BRD）是一种4-羟基香豆素的取代衍生物，具有抗凝血活性和急性毒性。本文报道了利用溴敌隆的内源荧光发射特性，研究了溴敌隆与血浆蛋白牛血清白蛋白（BSA）和人血清白蛋白（HSA）的相互作用。溴敌隆在水性缓冲介质中发出微弱荧光，发射波长为397 nm。溴敌隆与血清白蛋白（SA）的结合导致荧光发射强度和稳态荧光各向异性（r（ss））的显著增强，伴随着10 nm的蓝移。在血清白蛋白-溴敌隆复合物中，色氨酸（Trp）残基的选择性激发导致从溴敌隆发射，从而指示从Trp到BRD的福斯特型能量转移。BRD对Trp荧光的淬灭用于估计SA-BRD复合物的结合常数。BRD与BSA和HSA的结合常数分别为7.5 × 10（4）和3.7 × 10（5）L mol（-1）。在此基础上，提出了一种以水杨酸为荧光增强剂测定水样中BRD的新方法。溴敌隆的检测限在最佳条件下分别为0.77和0.19 μ g mL（-1）在BSA和HSA的存在下，分别。(c)2005 Elsevier B. V.保留所有权利。
Bromadiolone (BRD), a substituted 4-hydroxycoumarin derivative, is known to possess anti-coagulant activity with acute toxicity. In this paper, we report a study on the interaction of bromadiolone with the plasma proteins bovine serum albumin (BSA) and human serum albumin (HSA), using the intrinsic fluorescence emission properties of bromadiolone. Bromadiolone is weakly fluorescent in aqueous buffer medium, with an emission at 397 nm. Binding of bromadiolone with serum albumins (SA) leads to a marked enhancement in the fluorescence emission intensity and steady state fluorescence anisotropy (r(ss)) accompanied by a blueshift of 10 nm. In the serum albumin-bromadiolone complex, selective excitation of tryptophan (Trp) residue results in emission from bromadiolone, thereby indicating a Forster type energy transfer from Trp to BRD. This quenching of Trp fluorescence by BRD was used to estimate the binding constant of the SA-BRD complex. The binding constants for BRD with BSA and HSA were 7.5 x 10(4) and 3.7 x 10(5) L mol(-1), respectively. Based on this, a new method involving SA as fluorescence-enhancing reagent for estimation of BRD in aqueous samples has been suggested. The detection limits of bromadiolone under the optimum conditions were 0.77 and 0.19 mu g mL(-1) in presence of BSA and HSA, respectively. (c) 2005 Elsevier B.V. All rights reserved.