Marin Egor, Kovalev Kirill, Poelman Therese, Veenstra Rick, Borshchevskiy Valentin, Guskov Albert
Groningen Institute for Biomolecular Sciences and Biotechnology, University of Groningen, 9747AG Groningen, The Netherlands.
European Molecular Biology Laboratory, EMBL Hamburg c/o DESY, 22607 Hamburg, Germany.
Cryst Growth Des. 2023 Dec 12;24(1):325-330. doi: 10.1021/acs.cgd.3c01034. eCollection 2024 Jan 3.
Membrane proteins are indispensable for every living organism, yet their structural organization remains underexplored. Despite the recent advancements in single-particle cryogenic electron microscopy and cryogenic electron tomography, which have significantly increased the structural coverage of membrane proteins across various kingdoms, certain scientific methods, such as time-resolved crystallography, still mostly rely on crystallization techniques, such as lipidic cubic phase (LCP) or crystallization. In this study, we present an open-access blueprint for a humidity control chamber designed for LCP/ crystallization experiments using a Gryphon crystallization robot. Using this chamber, we have obtained crystals of a transmembrane aspartate transporter Glt from in a lipidic environment using crystallization. The data collected from these crystals allowed us to perform an analysis of lipids bound to the transporter. With this publication of our open-access design of a humidity chamber, we aim to improve the accessibility of protein crystallization for the scientific community.
膜蛋白对每个生物体来说都是不可或缺的，然而它们的结构组织仍未得到充分探索。尽管单颗粒低温电子显微镜和低温电子断层扫描技术最近取得了进展，显著增加了跨不同生物界的膜蛋白的结构覆盖范围，但某些科学方法，如时间分辨晶体学，仍然主要依赖于脂质立方相（LCP）或结晶等结晶技术。在本研究中，我们展示了一个用于使用Gryphon结晶机器人进行LCP/结晶实验的湿度控制室的开放获取蓝图。使用这个控制室，我们在脂质环境中通过结晶获得了来自 的跨膜天冬氨酸转运蛋白Glt的晶体。从这些晶体收集的数据使我们能够对与转运蛋白结合的脂质进行分析。通过发表我们湿度控制室的开放获取设计，我们旨在提高科学界对 蛋白质结晶的可及性。
