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首页> 美国卫生研究院文献>Acta Crystallographica Section F: Structural Biology and Crystallization Communications >Efficient UV detection of protein crystals enabled by fluorescence excitation at wavelengths longer than 300 nm
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Efficient UV detection of protein crystals enabled by fluorescence excitation at wavelengths longer than 300 nm

机译：通过波长大于300 nm的荧光激发可以对蛋白质晶体进行高效的紫外线检测

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摘要

It is well known that most proteins and many other biomolecules fluoresce when illuminated with UV radiation, but it is also commonly accepted that utilizing this property to detect protein crystals in crystallization setups is limited by the opacity of the materials used to contain and seal them. For proteins, this fluorescence property arises primarily from the presence of tryptophan residues in the sequence. Studies of protein crystallization results in a variety of setup configurations show that the opacity of the containment hardware can be overcome at longer excitation wavelengths, where typical hardware materials are more transparent in the UV, by the use of a powerful UV-light source that is effective in excitation even though not at the maximum of the excitation response. The results show that under these circumstances UV evaluation of crystallization trials and detection of biomolecular crystals in them is not limited by the hardware used. It is similarly true that a deficiency in tryptophan or another fluorescent component that limits the use of UV light for these purposes can be effectively overcome by the addition of fluorescent prostheses that bind to the biomolecule under study. The measurements for these studies were made with a device consisting of a potent UV-light source and a detection system specially adapted (i) to be tunable via a motorized and software-controlled absorption-filter system and (ii) to convey the excitation light to the droplet or capillary hosting the crystallization experiment by quartz-fibre light guides.
机译：众所周知，大多数蛋白质和许多其他生物分子在受到紫外线照射时会发出荧光，但也被普遍接受的是，利用此特性检测结晶设置中的蛋白质晶体受到用于容纳和密封它们的材料的不透明性的限制。对于蛋白质，这种荧光特性主要是由于序列中存在色氨酸残基而引起的。对蛋白质结晶结果进行各种设置的研究表明，通过使用功能强大的紫外线光源，可以在更长的激发波长下克服安全壳硬件的不透明性，在更长的激发波长下，典型的硬件材料在UV中更透明。即使不是最大的激励响应，也能有效激励。结果表明，在这种情况下，对结晶试验的UV评估和其中的生物分子晶体的检测不受所用硬件的限制。类似地，可以通过添加与研究中的生物分子结合的荧光假体来有效地克服色氨酸或其他荧光成分的不足，这些不足限制了将紫外线用于这些目的。这些研究的测量是使用由强大的紫外线光源和检测系统组成的设备进行的，该设备特别适合（i）通过电动和软件控制的吸收过滤器系统进行调谐，以及（ii）传输激发光通过石英纤维光导进入液滴或毛细管进行结晶实验。

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