Crystallographic studies of structures and mechanisms of dioxygen-employing enzymes or oxygen-sensitive proteins usually require these molecules to be prepared and crystallized under anoxic conditions in gloveboxes. Handling, cryo-cooling and extraction of these crystals from gloveboxes for X-ray data collection can be a tricky operation, during which samples might be ruined if they are accidentally exposed to atmospheric oxygen. This work reports the design of a new miniature airlock system and describes the associated method in order to simplify these tasks. The system allows flash-cooling of crystals harvested in a glovebox directly into an external liquid nitrogen container via a miniature airlock while maintaining strict anoxic conditions. The method was assessed using hen egg-white lysozyme crystals soaked in dihydroresorufin (an O₂ indicator) and with the reduced tRNA-hydroxylase enzyme MiaE.

中文翻译：

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一种微型气闸系统，有助于对在缺氧条件下生长的蛋白质晶体进行低温冷却

使用分子氧的酶或氧敏感蛋白的结构和机制的晶体学研究通常需要在手套箱中的缺氧条件下制备和结晶这些分子。从手套箱中处理、低温冷却和提取这些晶体以收集 X 射线数据可能是一项棘手的操作，在此过程中，如果样品意外暴露在大气中的氧气中，可能会损坏。这项工作报告了一种新的微型气闸系统的设计，并描述了相关的方法以简化这些任务。该系统允许将手套箱中收获的晶体通过微型气闸直接快速冷却到外部液氮容器中，同时保持严格的缺氧条件。该方法使用浸泡在二氢试卤灵（一种 O₂指标）和还原的tRNA-羟化酶MiaE。