Castellano F N, Dattelbaum J D, Lakowicz J R
Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore 21201, USA.
Anal Biochem. 1998 Jan 15;255(2):165-70. doi: 10.1006/abio.1997.2468.
We report the synthesis and spectral properties of two long-lifetime highly luminescent Ru(II) complexes containing either a sulfhydryl reactive iodoacetamido group or a less reactive choloroacetamido group, [Ru(bpy)2(5-iodoacetamido-1,10-phenanthroline)] (PF6)2 and Ru(bpy)2(5-chloroacetamido-1,10-phenanthroline) 2, respectively, where bpy is 2,2'-bipyridine. Ru(bpy)2(phen-IA)](PF6)2 was covalently linked to human serum albumin (HSA) and human immunoglobulin G (IgG). The photoluminescence lifetime of protein-bound probes approaches 1 microsecond under ambient conditions. In the absence of rotational motions, this probe displayed an anisotropy of 0.18 for excitation at 472 nm. Anisotropy decay data were used to determine the overall rotational correlation times of HSA and IgG. These long-lifetime sulfhydryl-reactive probes can be used to recover microsecond rotational motions and/or domain motions of proteins and/or macromolecular complexes.
我们报告了两种长寿命高发光钌（II）配合物的合成及光谱性质，这两种配合物分别含有巯基反应性碘乙酰胺基或反应性较低的氯乙酰胺基，即Ru(bpy)2(5 - 碘乙酰胺基 - 1,10 - 菲咯啉)2和Ru(bpy)2(5 - 氯乙酰胺基 - 1,10 - 菲咯啉)2，其中bpy为2,2'-联吡啶。Ru(bpy)2(phen - IA)](PF6)2与人类血清白蛋白（HSA）和人类免疫球蛋白G（IgG）共价连接。在环境条件下，与蛋白质结合的探针的光致发光寿命接近1微秒。在没有旋转运动的情况下，该探针在472 nm激发时显示出0.18的各向异性。各向异性衰减数据用于确定HSA和IgG的整体旋转相关时间。这些长寿命的巯基反应性探针可用于恢复蛋白质和/或大分子复合物的微秒级旋转运动和/或结构域运动。
