A simple and versatile microfluidic device for efficient biomacromolecule crystallization and structural analysis by serial crystallography

IUCrJ

Roche, J.¹  de Wijn, R.²  Hennig, O.³  Betat, H.⁴  Fernandez-Millan, P.⁵  Brillet, K.⁶  Rollet, K.⁷  Engilberge, S.⁸ 

[1] Architecture et Fonction des Macromolcules Biologiques, UMR 7257 CNRSAix Marseille University, 163 Avenue de Luminy, 13288 Marseille, France;Architecture et Ractivit de l'ARN, UPR 9002, CNRS, Institut de Biologie Molculaire et Cellulaire (IBMC), Universit de Strasbourg, 15 Rue Ren Descartes, 67084 Strasbourg, France;Institute for Biochemistry, Leipzig University, Bruederstrasse 34, 04103 Leipzig, Germany;Laboratorio de Estudios Cristalogrficos, IACT, CSICUniversidad de Granada, Avenida Las Palmeras 4, 18100 Armilla, Granada, Spain;PROXIMA 2A beamline, Synchrotron SOLEIL, L'Orme des Merisiers, Saint-Aubin, 91192 Gif-sur-Yvette, France;Paul Scherrer Institute, Swiss Light Source, Forschungsstrasse 111, 5232 Villigen PSI, Switzerland;Structural Biology, European Synchrotron Radiation Facility, 38043 Grenoble, France;Universit Grenoble Alpes, CEA, CNRS, IBS, 38000 Grenoble, France

关键词: MACROMOLECULE;    CRYSTALLIZATION;    COUNTER-DIFFUSION;    MICROFLUIDICS;    SEEDING;    LIGAND SOAKING;    TRACE FLUORESCENT LABELING;    SERIAL CRYSTALLOGRAPHY;    ROOM TEMPERATURE;    PROTEIN STRUCTURE;    CHIPX3;   

DOI  :  10.1107/S2052252519003622

学科分类：数学（综合）

来源: International Union of Crystallography

【 摘 要 】

Determining optimal conditions for the production of well diffracting crystals is a key step in every biocrystallography project. Here, a microfluidic device is described that enables the production of crystals by counter-diffusion and their direct on-chip analysis by serial crystallography at room temperature. Nine `non-model' and diverse biomacromolecules, including seven soluble proteins, a membrane protein and an RNA duplex, were crystallized and treated on-chip with a variety of standard techniques including micro-seeding, crystal soaking with ligands and crystal detection by fluorescence. Furthermore, the crystal structures of four proteins and an RNA were determined based on serial data collected on four synchrotron beamlines, demonstrating the general applicability of this multipurpose chip concept.

【 授权许可】

CC BY