| 八肋游仆虫中心蛋白N-端半分子与Tb3+，Ca2+的结合 | |
| Terbium- and Calcium-binding Properties of N-terminal Domain of Euplotes Centrin | |
| 摘要: 用分子生物学方法表达、纯化了游仆虫中心蛋白及N-端半分子，用铽荧光探针法、离子竞争法研究了pH 7.4，0.01 mol· L-1 Hepes条件下中心蛋白与铽、钙的结合性质。结果表明中心蛋白有4个铽结合部位，其中2个为高亲合结合部位、2个为低亲合结合部位。具有2个低亲合结合部位的中心蛋白半分子与铽结合的条件常数是(2.13±0.10)×105 L·mol-1，与钙结合的条件常数是(7.52±0.02)×102 L·mol-1。 | |
| 关键词: 中心蛋白 荧光 铽离子 钙离子 | |
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| Abstract: Euplotes centrins (EoCens) including full-length protein (apoEoCen) and semi-molecule centrin (N-apoEoCen) were expressed and purified by biological engineering. The binding of terbium (Tb3+) to apoEoCen was monitored by fluorescence in 0.01 mol·L-1 N-2-hydro-xyethylpiperazine-N-2-ethanesulfonic acid (Hepes), at pH 7.4. It could be seen that the protein binds two Tb3+ with high affinity and two Tb3+ with low affinity. The reactions between N-apoEoCen and Tb3+ or Ca2+ were also studied by Tb3+ fluorescence probe and ionic competition in 0.01 mol·L-1 Hepes, at pH 7.4. On the basis of fluorescence titration curves, the conditional binding constant of Tb2-N-EoCen was determined to be (2.13 ± 0.10) × 105 L·mol-1 and the relative binding constant of Ca2-N-EoCen was calculated to be (7.52 ± 0.02) × 102 L·mol-1. | |
| Keywords: Euplotes centrin fluorescence Tb3+ Ca2+ | |
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| 任列香,赵亚琴,丰九英,贺晓静,梁爱华,杨斌盛.八肋游仆虫中心蛋白N-端半分子与Tb3+，Ca2+的结合[J].无机化学学报,2006,22(1):87-90. | |
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