Takashi R
Biochemistry. 1979 Nov 13;18(23):5164-9. doi: 10.1021/bi00590a021.
The fast-reacting thiol (SH1) of myosin subfragment-1 (S-1) was covalently and specifically labeled with (iodoacetamido)fluorescein (IAF), while Cys-373 of actin was also covalently and preferentially labeled with N-(iodoacetyl)-N'-(1-sulfo-5-naphthyl)ethylenediamine (1,5-IAEDANS). The method of fluorescence energy transfer was used to examine the spatial proximity between the two sites, i.e., SH1 and Cys-373, in the rigor complex of acto-S-1. Approximately 30% fluorescence energy transfer was observed from the 1,5-IAEDANS on actin as a donor to the IAF on S-1 as an acceptor in their rigor complex; under certain assumptions this corresponds to a distance of ca. 6.0 nm.
肌球蛋白亚片段-1（S-1）的快速反应巯基（SH1）用（碘乙酰胺基）荧光素（IAF）进行了共价且特异性标记，而肌动蛋白的半胱氨酸-373也用N-（碘乙酰基）-N'-（1-磺基-5-萘基）乙二胺（1,5-IAEDANS）进行了共价且优先标记。荧光能量转移方法用于检测肌动蛋白-S-1的强直复合物中两个位点，即SH1和半胱氨酸-373之间的空间接近度。在它们的强直复合物中，观察到从肌动蛋白上作为供体的1,5-IAEDANS到S-1上作为受体的IAF约有30%的荧光能量转移；在某些假设下，这对应于约6.0纳米的距离。
