Miki M, Wahl P
Biochim Biophys Acta. 1984 Nov 9;790(3):275-83. doi: 10.1016/0167-4838(84)90032-3.
Fluorescence energy transfer was measured by time-resolved and steady-state fluorimetry in order to investigate the spatial relationships between the nucleotide binding site of actin, the Cys-373 residue of actin, and the SH1 of myosin subfragment-1 in the rigor complex of acto-subfragment-1. N-Iodoacetyl-N'-(5-sulfo-1-naphthyl)ethylenediamine (IAEDANS) bound to the Cys-373 of actin or the fluorescent ADP analogue 1-N6-ethenoadenosine-5'-diphosphate (epsilon-ADP) bound to F-actin was used as a donor and 4-(N-(iodoacetoxy)ethyl-N-methyl)amino-7-nitrobenz-2-oxa-1,3-diazo le (IANBD) or 5-iodoacetamidofluorescein (IAF) bound to SH1 of myosin subfragment-1 was used as an acceptor. Assuming the random orientation factor, K2, to be 2/3, the distance between Cys-373 residue of actin and SH1 of myosin subfragment-1 was calculated to be about 50 A, in agreement with the values previously reported, 60 A (Takashi, R. (1969) Biochemistry 18, 5164-69) and 50 A (Trayer, H.R. and Trayer, I.P. (1983) Eur. J. Biochem. 135, 47-59). The distance between the nucleotide binding site of actin and SH1 of myosin subfragment-1 was calculated to be about 70 A or greater.
为了研究肌动蛋白的核苷酸结合位点、肌动蛋白的半胱氨酸 - 373残基与肌动蛋白 - 肌球蛋白亚片段 - 1紧密复合物中肌球蛋白亚片段 - 1的SH1之间的空间关系，采用时间分辨荧光光谱法和稳态荧光光谱法测量了荧光能量转移。与肌动蛋白的半胱氨酸 - 373结合的N - 碘乙酰基 - N' -（5 - 磺基 - 1 - 萘基）乙二胺（IAEDANS）或与F - 肌动蛋白结合的荧光ADP类似物1 - N6 - 乙烯基腺苷 - 5'-二磷酸（ε - ADP）用作供体，与肌球蛋白亚片段 - 1的SH1结合的4 -（N -（碘乙酰氧基）乙基 - N - 甲基）氨基 - 7 - 硝基苯并 - 2 - 恶唑 - 1,3 - 二氮杂萘（IANBD）或5 - 碘乙酰氨基荧光素（IAF）用作受体。假设随机取向因子K2为2/3，计算得出肌动蛋白的半胱氨酸 - 373残基与肌球蛋白亚片段 - 1的SH1之间的距离约为50埃，这与先前报道的值一致，分别为60埃（Takashi，R.（1969年）《生物化学》18, 5164 - 69）和50埃（Trayer，H.R.和Trayer，I.P.（1983年）《欧洲生物化学杂志》135, 47 - 59）。计算得出肌动蛋白的核苷酸结合位点与肌球蛋白亚片段 - 1的SH1之间的距离约为70埃或更大。
