Photophysical properties and synthesis of new dye-cyclooctyne conjugates for multicolor and advanced microscopy.
Photophysical properties and synthesis of new dye-cyclooctyne conjugates for multicolor and advanced microscopy.
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用于多色和高级显微镜的新型染料-环辛炔缀合物的光物理性质和合成。
DOI：
10.1021/acs.bioconjchem.5b00059
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发表时间：
2015
影响因子：
4.7
通讯作者：
Stefan Bräse
中科院分区：
文献类型：
--
作者：
A. Hörner;Tobias Hagendorn;U. Schepers;Stefan Bräse
具有荧光团的环辛炔缀合物通常用于细胞和组织中的生物正交标记。然而，迄今为止还没有描述具有跨越整个可见光谱直到NIR的不同荧光染料的一个环辛炔核心结构的综合库。因此，我们合成并评价了一种环辛炔核心结构，该结构易于连接不同的染料用于荧光成像、FRET分析和体内代谢研究。由于这些原因，我们开发了一个简单的一步合成从已知的环辛炔。与NHS活化的染料组合，环辛炔在室温下仅在1-2小时内以高产率与染料DAB-MFCO缀合物反应。我们用具有高亮度并且在从绿色到NIR的波长下漂白稳定的染料创建了缀合物。测试了标记细胞表面上聚糖的能力。所有染料DAB-MFCO缀合物都在叠氮官能化聚糖结构上进行点击反应，具有令人满意的光物理性质。总共合成了七种不同的染料DAB-MFCO缀合物;评估了它们的生物物理性质和在生物应用中用于点击标记的适用性，使它们适合于单分子和高分辨率测量。
Cyclooctyne conjugates with fluorophores are often used for bioorthogonal labeling in cells and tissues. However, no comprehensive library of one cyclooctyne core structure with different fluorescent dyes spanning the whole visible spectrum up to the NIR had been described so far. Hence, we synthesized and evaluated one cyclooctyne core structure which is easily accessible for the attachment of different dyes for multicolor imaging, FRET analysis, and study of metabolism in vivo. For these reasons we developed an easy one step synthesis starting from a known cyclooctyne. In combination with NHS-activated dyes, the cyclooctyne reacted to the dye DAB-MFCO conjugates within only 1-2 h at room temperature with high yields. We created conjugates with dyes that have high brightness and are bleaching stable with wavelengths from green to NIR. The ability to label glycans on cell surfaces was tested. All dye DAB-MFCO conjugates undergo click reactions on azide functionalized glycan structures with satisfactory photophysical properties. In total, seven different dye DAB-MFCO conjugates were synthesized; their photophysical properties and suitability for click labeling in biological applications were evaluated, making them suitable for single molecule and high resolution measurements.
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影响因子：
16.6
作者：
Ning, Xinghai;Guo, Jun;Boons, Geert-Jan
通讯作者：
Boons, Geert-Jan
影响因子：
15
作者：
John T. Wilson;Venkata R. Krishnamurthy;W. Cui;Z. Qu;E. Chaikof
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John T. Wilson;Venkata R. Krishnamurthy;W. Cui;Z. Qu;E. Chaikof
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Hein JE;Fokin VV
通讯作者：
Fokin VV
影响因子：
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作者：
Agard, NJ;Prescher, JA;Bertozzi, CR
通讯作者：
Bertozzi, CR
影响因子：
16.6
作者：
Sletten, Ellen M.;Bertozzi, Carolyn R.
通讯作者：
Bertozzi, Carolyn R.