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bioRxiv · 10.1101/2021.06.03.446146

An automated platform for structural analysis of membrane proteins through serial crystallography

Healey, R. D.·Basu, S.·Humm, A.-S.·Leyrat, C.·Cong, X.·Golebiowski, J.·Dupeux, F.·Pica, A.·Granier, S.·Marquez, J. A.
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Abstract

Membrane proteins are central to many pathophysiological processes yet remain very difficult to analyze at a structural level. Moreover, high-throughput structure-based drug discovery has not yet been exploited for membrane proteins due to lack of automation. Here, we present a facile and versatile platform for in meso membrane protein crystallization, enabling rapid atomic structure determination at both cryogenic and room temperature and in a single support. We apply this approach to two human integral membrane proteins, which allowed us to capture different conformational states of intramembrane enzyme-product complexes and analyze the structural dynamics of the ADIPOR2 integral membrane protein. Finally, we demonstrate an automated pipeline combining high-throughput microcrystal soaking, automated laser-based harvesting and serial crystallography enabling screening of small molecule libraries with membrane protein crystals grown in meso. This approach brings badly needed automation for this important class of drug targets and enables high-throughput structure-based ligand discovery with membrane proteins. HighlightsO_LIA fully automated, online workflow enables rapid determination of membrane protein structures by serial X-ray crystallography (SSX). C_LIO_LIHigh resolution room temperature and cryogenic structures of ADIPOR2 provide insights into the dynamic nature of receptor:ligand interactions. C_LIO_LIA web-based application allows remote user-guided experimental design and execution. C_LIO_LIAn automated SSX-based ligand discovery pipeline for integral membrane proteins is introduced. C_LI

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Healey, R. D., Basu, S., Humm, A.-S., Leyrat, C., Cong, X., Golebiowski, J., Dupeux, F., Pica, A., Granier, S., Marquez, J. A.. 2021-06-03. An automated platform for structural analysis of membrane proteins through serial crystallography. https://doi.org/10.1101/2021.06.03.446146

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