Multicomponent mixtures for cryoprotection and ligand solubilization
Mixed cryoprotectants have been developed for the solubilization of ligands for crystallization of protein–ligand complexes and for crystal soaking. Low affinity lead compounds with poor solubility are problematic for structural studies. Complete ligand solubilization is required for co-crystallizat...
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doaj-89ef1af888194602a35159ce0497cba32020-11-25T00:33:01ZengElsevierBiotechnology Reports2215-017X2015-09-017C12012710.1016/j.btre.2015.05.008Multicomponent mixtures for cryoprotection and ligand solubilizationLidia Ciccone0Laura Vera1Livia Tepshi2Lea Rosalia3Armando Rossello4Enrico A. Stura5CEA, iBiTec-S, Service d’Ingénierie Moléculaire des Protéines, Laboratoire de Toxinologie Moléculaire et Biotechnologies, Gif-sur-Yvette, F-91191, FranceCEA, iBiTec-S, Service d’Ingénierie Moléculaire des Protéines, Laboratoire de Toxinologie Moléculaire et Biotechnologies, Gif-sur-Yvette, F-91191, FranceCEA, iBiTec-S, Service d’Ingénierie Moléculaire des Protéines, Laboratoire de Toxinologie Moléculaire et Biotechnologies, Gif-sur-Yvette, F-91191, FranceCEA, iBiTec-S, Service d’Ingénierie Moléculaire des Protéines, Laboratoire de Toxinologie Moléculaire et Biotechnologies, Gif-sur-Yvette, F-91191, FranceDipartimento di Farmacia, Università di Pisa, Via Bonanno 6, 56126 Pisa, ItalyCEA, iBiTec-S, Service d’Ingénierie Moléculaire des Protéines, Laboratoire de Toxinologie Moléculaire et Biotechnologies, Gif-sur-Yvette, F-91191, FranceMixed cryoprotectants have been developed for the solubilization of ligands for crystallization of protein–ligand complexes and for crystal soaking. Low affinity lead compounds with poor solubility are problematic for structural studies. Complete ligand solubilization is required for co-crystallization and crystal soaking experiments to obtain interpretable electron density maps for the ligand. Mixed cryo-preserving compounds are needed prior to X-ray data collection to reduce radiation damage at synchrotron sources. Here we present dual-use mixes that act as cryoprotectants and also promote the aqueous solubility of hydrophobic ligands. Unlike glycerol that increases protein solubility and can cause crystal melting the mixed solutions of cryo-preserving compounds that include precipitants and solubilizers, allow for worry-free crystal preservation while simultaneously solubilizing relatively hydrophobic ligands, typical of ligands obtained in high-throughput screening. The effectiveness of these mixture has been confirmed on a human transthyretin crystals both during crystallization and in flash freezing of crystals.http://www.sciencedirect.com/science/article/pii/S2215017X15000351 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Lidia Ciccone Laura Vera Livia Tepshi Lea Rosalia Armando Rossello Enrico A. Stura |
spellingShingle |
Lidia Ciccone Laura Vera Livia Tepshi Lea Rosalia Armando Rossello Enrico A. Stura Multicomponent mixtures for cryoprotection and ligand solubilization Biotechnology Reports |
author_facet |
Lidia Ciccone Laura Vera Livia Tepshi Lea Rosalia Armando Rossello Enrico A. Stura |
author_sort |
Lidia Ciccone |
title |
Multicomponent mixtures for cryoprotection and ligand solubilization |
title_short |
Multicomponent mixtures for cryoprotection and ligand solubilization |
title_full |
Multicomponent mixtures for cryoprotection and ligand solubilization |
title_fullStr |
Multicomponent mixtures for cryoprotection and ligand solubilization |
title_full_unstemmed |
Multicomponent mixtures for cryoprotection and ligand solubilization |
title_sort |
multicomponent mixtures for cryoprotection and ligand solubilization |
publisher |
Elsevier |
series |
Biotechnology Reports |
issn |
2215-017X |
publishDate |
2015-09-01 |
description |
Mixed cryoprotectants have been developed for the solubilization of ligands for crystallization of protein–ligand complexes and for crystal soaking. Low affinity lead compounds with poor solubility are problematic for structural studies. Complete ligand solubilization is required for co-crystallization and crystal soaking experiments to obtain interpretable electron density maps for the ligand. Mixed cryo-preserving compounds are needed prior to X-ray data collection to reduce radiation damage at synchrotron sources. Here we present dual-use mixes that act as cryoprotectants and also promote the aqueous solubility of hydrophobic ligands. Unlike glycerol that increases protein solubility and can cause crystal melting the mixed solutions of cryo-preserving compounds that include precipitants and solubilizers, allow for worry-free crystal preservation while simultaneously solubilizing relatively hydrophobic ligands, typical of ligands obtained in high-throughput screening. The effectiveness of these mixture has been confirmed on a human transthyretin crystals both during crystallization and in flash freezing of crystals. |
url |
http://www.sciencedirect.com/science/article/pii/S2215017X15000351 |
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