Orlans Julien, Rose Samuel L, Ferguson Gavin, Oscarsson Marcus, Homs Puron Alejandro, Beteva Antonia, Debionne Samuel, Theveneau Pascal, Coquelle Nicolas, Kieffer Jerome, Busca Paolo, Sinoir Jeremy, Armijo Victor, Lopez Marrero Marcos, Felisaz Franck, Papp Gergely, Gonzalez Herve, Caserotto Hugo, Dobias Fabien, Gigmes Jonathan, Lebon Guillaume, Basu Shibom, de Sanctis Daniele
ESRF - The European Synchrotron, 71 Avenue des Martyrs, Grenoble, France.
Institut de Génomique Fonctionnelle, Université de Montpellier, CNRS, INSERM, Montpellier, France.
Commun Chem. 2025 Jan 7;8(1):6. doi: 10.1038/s42004-024-01404-y.
Serial macromolecular crystallography has become a powerful method to reveal room temperature structures of biological macromolecules and perform time-resolved studies. ID29, a flagship beamline of the ESRF 4th generation synchrotron, is the first synchrotron beamline in the world capable of delivering high brilliance microsecond X-ray pulses at high repetition rate for the structure determination of biological macromolecules at room temperature. The cardinal combination of microsecond exposure times, innovative beam characteristics and adaptable sample environment provides high quality complete data, even from an exceptionally small amount of crystalline material, enabling what we collectively term serial microsecond crystallography (SµX). After validating the use of different sample delivery methods with various model systems, we applied SµX to an integral membrane receptor, where only a few thousands diffraction images were sufficient to obtain a fully interpretable electron density map for the antagonist istradefylline-bound A receptor conformation, providing access to the antagonist binding mode. SµX, as demonstrated at ID29, will quickly find its broad applicability at upcoming 4th generation synchrotron sources worldwide and opens a new frontier in time-resolved SµX.
串行大分子晶体学已成为揭示生物大分子室温结构并进行时间分辨研究的强大方法。ID29是欧洲同步辐射装置（ESRF）第四代同步加速器的旗舰光束线，是世界上第一条能够以高重复率提供高亮度微秒级X射线脉冲，用于在室温下测定生物大分子结构的同步加速器光束线。微秒级曝光时间、创新的光束特性和适应性强的样品环境的关键组合，即使从极少量的晶体材料中也能提供高质量的完整数据，从而实现了我们统称为串行微秒晶体学（SµX）的技术。在用各种模型系统验证了不同样品输送方法的使用后，我们将SµX应用于一种整合膜受体，在该受体中，仅几千张衍射图像就足以获得用于拮抗剂异丁司特结合的A受体构象的完全可解释的电子密度图，从而揭示拮抗剂的结合模式。如在ID29上所展示的，SµX将很快在全球即将建成的第四代同步加速器光源中找到广泛的应用，并开启时间分辨SµX的新前沿。
