Metal-ligand complexes as a new class of long-lived fluorophores for protein hydrodynamics.
Metal-ligand complexes as a new class of long-lived fluorophores for protein hydrodynamics.
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DOI：
10.1016/s0006-3495(95)80193-1
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发表时间：
1995
影响因子：
3.4
通讯作者：
Ewald Terpetschnig;H. Szmacinski;H. Malak;J. Lakowicz
中科院分区：
文献类型：
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作者：
Ewald Terpetschnig;H. Szmacinski;H. Malak;J. Lakowicz
我们描述了使用的不对称钌配体配合物作为一类新的发光探针，可用于测量旋转运动的蛋白质。已知这些配合物显示10至4000 ns的发光寿命。在这份报告中，我们表明，不对称配合物Ru（bpy）2（dcbpy）（PF 6）2显示出高的各向异性值时，在长波长的吸收带激发。为了与蛋白质共价连接，我们合成了这种金属配体络合物的N-羟基琥珀酰亚胺酯。为了说明这些探针的有用性，我们描述了[Ru（bpy）2（dcbpy）]的强度和各向异性衰减时，共价连接到人血清白蛋白，刀豆球蛋白A（ConA），人免疫球蛋白G（IgG），和铁蛋白，并在增加粘度的溶液中测量。这些数据表明，探针可用于测量旋转运动的10 ns至1.5微秒的时间尺度，这是迄今为止一直无法使用发光方法。本探针[Ru（bpy）2（dcbpy）]可被视为一类金属配体配合物中的第一个，每个配合物具有不同的化学反应性和光谱性质，用于大分子动力学的研究。
We describe the use of asymmetric Ru-ligand complexes as a new class of luminescent probes that can be used to measure rotational motions of proteins. These complexes are known to display luminescent lifetimes ranging from 10 to 4000 ns. In this report, we show that the asymmetric complex Ru(bpy)2(dcbpy) (PF6)2 displays a high anisotropy value when excited in the long wavelength absorption band. For covalent linkage to proteins, we synthesized the N-hydroxy succinimide ester of this metal-ligand complex. To illustrate the usefulness of these probes, we describe the intensity and anisotropy decays of [Ru(bpy)2(dcbpy)] when covalently linked to human serum albumin, concanavalin A (ConA), human immunoglobulin G (IgG), and Ferritin, and measured in solutions of increased viscosity. These data demonstrate that the probes can be used to measure rotational motions on the 10 ns to 1.5 microseconds timescale, which so far has been inaccessible using luminescence methods. The present probe [Ru(bpy)2(dcbpy)] can be regarded as the first of a class of metal-ligand complexes, each with different chemical reactivity and spectral properties, for studies of macromolecular dynamics.