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通过荧光发射光谱、微量热等温滴定法和荧光共振光散射方法研究了Tb~(3+)与磷酸化人中心蛋白1(phosphorylated human centrin 1,HsCen1p)的作用。结果表明,HsCen1p与Tb~(3+)的结合比为1∶4,与Ⅰ、Ⅱ、Ⅲ和Ⅳ位点的结合能力分别为(0.75±0.40)×10~4,(0.70±0.40)×10~4,(8.50±0.40)×10~4和(1.23±0.07)×10~5(mol/L)~(-1),其结合顺序为Ⅳ>Ⅲ>Ⅰ>Ⅱ。另外,研究了Tb~(3+)对HsCen1p聚集性质的影响,发现蛋白HsCen1p的N端结构域在聚集过程中发挥重要作用。
The effects of Tb 3+ and phosphorylated human centrin 1 (HsCen1p) were studied by fluorescence emission spectroscopy, microthermal thermal isothermal titration and fluorescence resonance light scattering. The results showed that the binding ratio of HsCen1p to Tb ~ (3+) was 1: 4, and the binding ability of HsCen1p to (Ⅰ), Ⅱ, Ⅲ and Ⅳ were (0.75 ± 0.40) × 10 ~ 4 and (0.70 ± 0.40) × 10 ~ 4, (8.50 ± 0.40) × 10 ~ 4 and (1.23 ± 0.07) × 10 ~ 5 (mol / L) -1, respectively. The order of binding was Ⅳ> Ⅲ> Ⅰ> Ⅱ. In addition, the effect of Tb ~ (3+) on the aggregation of HsCen1p was studied, and the N-terminal domain of HsCen1p was found to play an important role in the aggregation process.