Revealing protein structures: crystallization of protein‐ligand complexes – co‐crystallization and crystal soaking

Protein crystallogenesis represents a key step in X‐ray crystallography studies that employ co‐crystallization and ligand soaking for investigating ligand binding to proteins. Co‐crystallization is a method that enables the precise determination of binding positions, although it necessitates a signi...

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Main Authors: Barbora Kaščáková, Anna Koutská, Michaela Burdová, Petra Havlíčková, Ivana Kutá Smatanová
Format: Article
Language:English
Published: Wiley 2025-04-01
Series:FEBS Open Bio
Subjects:
Online Access:https://doi.org/10.1002/2211-5463.13913
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author Barbora Kaščáková
Anna Koutská
Michaela Burdová
Petra Havlíčková
Ivana Kutá Smatanová
author_facet Barbora Kaščáková
Anna Koutská
Michaela Burdová
Petra Havlíčková
Ivana Kutá Smatanová
author_sort Barbora Kaščáková
collection DOAJ
description Protein crystallogenesis represents a key step in X‐ray crystallography studies that employ co‐crystallization and ligand soaking for investigating ligand binding to proteins. Co‐crystallization is a method that enables the precise determination of binding positions, although it necessitates a significant degree of optimization. The utilization of microseeding can facilitate a reduction in sample requirements and accelerate the co‐crystallization process. Ligand soaking is the preferred method due to its simplicity; however, it requires careful control of soaking conditions to ensure the successful integration of the ligands. This research protocol details the procedures for co‐crystallization and soaking to achieve protein–ligand complex formation, which is essential for advancing drug discovery. Additionally, a simple protocol for demonstrating soaking for educational purposes is described.
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series FEBS Open Bio
spelling doaj-art-e92cf4107b864c849b4f2a35626bb93d2025-08-20T01:55:37ZengWileyFEBS Open Bio2211-54632025-04-0115454255010.1002/2211-5463.13913Revealing protein structures: crystallization of protein‐ligand complexes – co‐crystallization and crystal soakingBarbora Kaščáková0Anna Koutská1Michaela Burdová2Petra Havlíčková3Ivana Kutá Smatanová4Department of Chemistry, Faculty of Science University of South Bohemia in České Budějovice Czech RepublicDepartment of Chemistry, Faculty of Science University of South Bohemia in České Budějovice Czech RepublicDepartment of Chemistry, Faculty of Science University of South Bohemia in České Budějovice Czech RepublicDepartment of Chemistry, Faculty of Science University of South Bohemia in České Budějovice Czech RepublicDepartment of Chemistry, Faculty of Science University of South Bohemia in České Budějovice Czech RepublicProtein crystallogenesis represents a key step in X‐ray crystallography studies that employ co‐crystallization and ligand soaking for investigating ligand binding to proteins. Co‐crystallization is a method that enables the precise determination of binding positions, although it necessitates a significant degree of optimization. The utilization of microseeding can facilitate a reduction in sample requirements and accelerate the co‐crystallization process. Ligand soaking is the preferred method due to its simplicity; however, it requires careful control of soaking conditions to ensure the successful integration of the ligands. This research protocol details the procedures for co‐crystallization and soaking to achieve protein–ligand complex formation, which is essential for advancing drug discovery. Additionally, a simple protocol for demonstrating soaking for educational purposes is described.https://doi.org/10.1002/2211-5463.13913advanced crystallizationco‐crystallizationcrystal soakingcrystallization protocolmicroseeding
spellingShingle Barbora Kaščáková
Anna Koutská
Michaela Burdová
Petra Havlíčková
Ivana Kutá Smatanová
Revealing protein structures: crystallization of protein‐ligand complexes – co‐crystallization and crystal soaking
FEBS Open Bio
advanced crystallization
co‐crystallization
crystal soaking
crystallization protocol
microseeding
title Revealing protein structures: crystallization of protein‐ligand complexes – co‐crystallization and crystal soaking
title_full Revealing protein structures: crystallization of protein‐ligand complexes – co‐crystallization and crystal soaking
title_fullStr Revealing protein structures: crystallization of protein‐ligand complexes – co‐crystallization and crystal soaking
title_full_unstemmed Revealing protein structures: crystallization of protein‐ligand complexes – co‐crystallization and crystal soaking
title_short Revealing protein structures: crystallization of protein‐ligand complexes – co‐crystallization and crystal soaking
title_sort revealing protein structures crystallization of protein ligand complexes co crystallization and crystal soaking
topic advanced crystallization
co‐crystallization
crystal soaking
crystallization protocol
microseeding
url https://doi.org/10.1002/2211-5463.13913
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