A glycine spacer improved peptidyl-nuclear-localized efficiency for fluorescent imaging nuclear H2O2
A glycine spacer improved peptidyl-nuclear-localized efficiency for fluorescent imaging nuclear H2O2
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甘氨酸间隔基提高了核 H2O2 荧光成像的肽基核定位效率
DOI：
10.1016/j.snb.2019.126624
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发表时间：
2019-10
期刊：
影响因子：
--
通讯作者：
Caixia Yin
中科院分区：
文献类型：
--
作者：
Ying Wen;Fangjun Huo;Caixia Yin
异常产生的H_2O_2可攻击细胞核内的DNA，导致DNA氧化损伤，与许多严重的疾病有关。因此，监测核H2 O2水平至关重要。然而，绝大多数的荧光传感器不能定位在细胞核内。在本研究中，我们使用甘氨酸（最简单的氨基酸）作为间隔子来延长N50和NP 1之间的距离。采用无Cu 2+点击反应（通过插入二苯并环辛炔-马来酰亚胺）将两个部分原位连接。测试了三个间隔区序列：C、CGGGGG和CGGGGGGGGG，并产生了三个新的NLS肽（连接间隔区序列至N50）：pep 5、pep 6和pep 7。荧光成像结果表明，三种多肽的核靶向性均有所提高，尤其是pep 6。通过分子对接模拟计算出的与importin α5（importin的一个成员）的结合能为pep 6 <pep 5 <pep 7，与荧光成像结果一致。重要的是，由pep 6和NP 1组成的探针系统用于成像刺激后核H2 O2水平的增加。
H2O2generated aberrantly can attack DNA in nuclei and result in oxidative DNA damage, which has been associated with many serious diseases. Therefore, it is crucial to monitor nuclear H2O2levels. However, the vast majority of fluorescence sensors can not localize in the nucleus. In the present study, we empolyed glycine (the simplest amino acids) as a spacer to lengthen distance betweenN50andNP1. Cu2+-free click reaction (through inserting dibenzocyclooctyne-maleimide) was used to connect the two partsin situ. Three spacer sequences were tested: C, CGGGGG, and CGGGGGGGGGG, and three new NLS peptides (connection spacer sequence toN50) were generated:pep5,pep6andpep7. The fluorescence imaging results showed the nucleus-targeting efficancy of the three peptides, especiallypep6, were imporved. The calculated binding energies by molecular docking simulations with importin α5 (a member of importin) werepep6 < pep5 < pep7, consistent with the fluorescence imaging results. Importantly, the probe system consisting ofpep6andNP1was used for imaging the increase of nuclear H2O2level upon stimulation.
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