Time-resolved crystallography enabled the visualization of protein molecular motion during reaction. While light is commonly used to initiate reactions in time-resolved crystallography, only a small number of proteins can in fact be activated by light. However, many biological reactions can be triggered by the interaction of proteins with ligands. The sample delivery method presented here uses a mix-and-extrude approach based on 3D printed microchannels in conjunction with a micronozzle to study the dynamics of samples in viscous media that can be triggered by diffusive mixing. The device design allows for mixing of ligands and protein crystals in a time window of 2 to 20 seconds. The device characterization using a model system (fluorescence quenching of iq-mEmerald proteins by copper ions) demonstrated that ligand and protein crystals, each within the lipidic cubic phase, can be mixed efficiently. The potential use of this approach for time-resolved membrane protein crystallography to support in the development of new drugs is also discussed.

中文翻译：

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混合和挤出：高粘度样品注入到时间分辨蛋白质晶体学

时间分辨晶体学使反应过程中蛋白质分子运动的可视化成为可能。虽然光通常用于启动时间分辨晶体学中的反应，但实际上只有少数蛋白质可以被光激活。然而，许多生物反应可以由蛋白质与配体的相互作用引发。此处介绍的样品输送方法使用基于 3D 打印微通道和微喷嘴的混合和挤出方法来研究粘性介质中可由扩散混合触发的样品动力学。该设备设计允许在 2 到 20 秒的时间窗口内混合配体和蛋白质晶体。使用模型系统（铜离子对 iq-mEmerald 蛋白的荧光淬灭）的器件表征表明，配体和蛋白质晶体，每个都在脂质立方相内，可以有效混合。还讨论了这种方法在时间分辨膜蛋白晶体学中用于支持新药开发的潜在用途。