Novel Device and Strategy for Growing Large, High-Quality Protein Crystals by Controlling Crystallization Conditions

Neutron diffraction experiments are informative for determining the locations of hydrogen atoms in protein molecules; however, much larger crystals are needed than those required for X-ray diffraction. Thus, additional techniques are required to grow larger crystals. Here, a unique crystallization d...

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Autores principales:	Naoki Tanigawa, Sachiko Takahashi, Bin Yan, Masayuki Kamo, Naoki Furubayashi, Koji Kubota, Koji Inaka, Hiroaki Tanaka
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	neutron diffraction large crystal reservoir control real-time observation Crystallography QD901-999
Acceso en línea:	https://doaj.org/article/aea33564518b4825970c771111e3b5fc
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spelling	oai:doaj.org-article:aea33564518b4825970c771111e3b5fc2021-11-25T17:18:07ZNovel Device and Strategy for Growing Large, High-Quality Protein Crystals by Controlling Crystallization Conditions10.3390/cryst111113112073-4352https://doaj.org/article/aea33564518b4825970c771111e3b5fc2021-10-01T00:00:00Zhttps://www.mdpi.com/2073-4352/11/11/1311https://doaj.org/toc/2073-4352Neutron diffraction experiments are informative for determining the locations of hydrogen atoms in protein molecules; however, much larger crystals are needed than those required for X-ray diffraction. Thus, additional techniques are required to grow larger crystals. Here, a unique crystallization device and strategy for growing large protein crystals are introduced. The device uses two micropumps to control crystal growth by altering the precipitant concentration and regulating the pinpoint injection of dry air flow to the crystallization cell. Furthermore, the crystal growth can be observed in real time. Preliminary microbatch crystallization experiments at various concentration ranges of polyethylene glycol (PEG) 4000 and sodium chloride were first performed to elucidate optimized crystallization conditions. Based on these results, a device to precisely control the sodium chloride and PEG concentrations and the supply of dry air to the crystallization cell was used, and 1.8 mm lysozyme and 1.5 mm alpha-amylase crystals with good reproducibility were obtained. X-ray data sets of both crystals were collected at room temperature at BL2S1 of the Aichi Synchrotron Radiation Center and confirmed that these crystals were of high quality. Therefore, this crystallization device and strategy were effective for growing large, high-quality protein crystals.Naoki TanigawaSachiko TakahashiBin YanMasayuki KamoNaoki FurubayashiKoji KubotaKoji InakaHiroaki TanakaMDPI AGarticleneutron diffractionlarge crystalreservoir controlreal-time observationCrystallographyQD901-999ENCrystals, Vol 11, Iss 1311, p 1311 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	neutron diffraction large crystal reservoir control real-time observation Crystallography QD901-999
spellingShingle	neutron diffraction large crystal reservoir control real-time observation Crystallography QD901-999 Naoki Tanigawa Sachiko Takahashi Bin Yan Masayuki Kamo Naoki Furubayashi Koji Kubota Koji Inaka Hiroaki Tanaka Novel Device and Strategy for Growing Large, High-Quality Protein Crystals by Controlling Crystallization Conditions
description	Neutron diffraction experiments are informative for determining the locations of hydrogen atoms in protein molecules; however, much larger crystals are needed than those required for X-ray diffraction. Thus, additional techniques are required to grow larger crystals. Here, a unique crystallization device and strategy for growing large protein crystals are introduced. The device uses two micropumps to control crystal growth by altering the precipitant concentration and regulating the pinpoint injection of dry air flow to the crystallization cell. Furthermore, the crystal growth can be observed in real time. Preliminary microbatch crystallization experiments at various concentration ranges of polyethylene glycol (PEG) 4000 and sodium chloride were first performed to elucidate optimized crystallization conditions. Based on these results, a device to precisely control the sodium chloride and PEG concentrations and the supply of dry air to the crystallization cell was used, and 1.8 mm lysozyme and 1.5 mm alpha-amylase crystals with good reproducibility were obtained. X-ray data sets of both crystals were collected at room temperature at BL2S1 of the Aichi Synchrotron Radiation Center and confirmed that these crystals were of high quality. Therefore, this crystallization device and strategy were effective for growing large, high-quality protein crystals.
format	article
author	Naoki Tanigawa Sachiko Takahashi Bin Yan Masayuki Kamo Naoki Furubayashi Koji Kubota Koji Inaka Hiroaki Tanaka
author_facet	Naoki Tanigawa Sachiko Takahashi Bin Yan Masayuki Kamo Naoki Furubayashi Koji Kubota Koji Inaka Hiroaki Tanaka
author_sort	Naoki Tanigawa
title	Novel Device and Strategy for Growing Large, High-Quality Protein Crystals by Controlling Crystallization Conditions
title_short	Novel Device and Strategy for Growing Large, High-Quality Protein Crystals by Controlling Crystallization Conditions
title_full	Novel Device and Strategy for Growing Large, High-Quality Protein Crystals by Controlling Crystallization Conditions
title_fullStr	Novel Device and Strategy for Growing Large, High-Quality Protein Crystals by Controlling Crystallization Conditions
title_full_unstemmed	Novel Device and Strategy for Growing Large, High-Quality Protein Crystals by Controlling Crystallization Conditions
title_sort	novel device and strategy for growing large, high-quality protein crystals by controlling crystallization conditions
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/aea33564518b4825970c771111e3b5fc
work_keys_str_mv	AT naokitanigawa noveldeviceandstrategyforgrowinglargehighqualityproteincrystalsbycontrollingcrystallizationconditions AT sachikotakahashi noveldeviceandstrategyforgrowinglargehighqualityproteincrystalsbycontrollingcrystallizationconditions AT binyan noveldeviceandstrategyforgrowinglargehighqualityproteincrystalsbycontrollingcrystallizationconditions AT masayukikamo noveldeviceandstrategyforgrowinglargehighqualityproteincrystalsbycontrollingcrystallizationconditions AT naokifurubayashi noveldeviceandstrategyforgrowinglargehighqualityproteincrystalsbycontrollingcrystallizationconditions AT kojikubota noveldeviceandstrategyforgrowinglargehighqualityproteincrystalsbycontrollingcrystallizationconditions AT kojiinaka noveldeviceandstrategyforgrowinglargehighqualityproteincrystalsbycontrollingcrystallizationconditions AT hiroakitanaka noveldeviceandstrategyforgrowinglargehighqualityproteincrystalsbycontrollingcrystallizationconditions
_version_	1718412552794275840

Novel Device and Strategy for Growing Large, High-Quality Protein Crystals by Controlling Crystallization Conditions

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