Optimization of Protein Crystallization: The OptiCryst Project

García-Caballero, Alfonso; Gavira, José A.; Pineda‐Molina, Estela; Chayen, Naomi E.; Govada, Lata; Khurshid, Sarfraz; Saridakis, Emmanuel; Boudjemline, Attia; Swann, Marcus J.; Stewart, Patrick D. Shaw; Briggs, Richard A.; Kolek, Stefan; Oberthüer, D.; Dierks, Karsten; Betzel, Christian; Santana, Martha; Hobbs, Jeanette R.; Thaw, Paul; Savill, Tony J.; Mesters, J.R.; Hilgenfeld, Rolf; Bonander, Nicklas; Bill, Roslyn M.

doi:10.1021/cg1013768

Cited by 15 publications

(20 citation statements)

References 49 publications

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“…For example, polymeric films containing ionizable groups, such as sulfonated polystyrene, have been tested as heterogeneous nucleation inducing surfaces for proteins, among other numerous materials and native and/or engineered surfaces [9]- [11]. Nucleation kinetics and thermodynamics studies indicated that significant conformational changes can occur as the protein interacts with these surfaces [10,12]. Recently, the use of nucleation agents to accelerate the crystallization of proteins was demonstrated [13].…”

Section: Introductionmentioning

confidence: 99%

“…In this study, we investigated crystallization of lysozyme (as a model protein) on three different metal surfaces to determine the optimal surfaces for the crystallization of proteins using the MA-MAEC technique and the iCrystal system (a mono-mode solid state microwave source with a variable power up to 100 W and a monomode microwave cavity with transverse electric mode, TE 10 ). The present study yielded the following advancements in protein crystallization: 1) the use of the MA-MAEC technique with the iCrystal system significantly reduces the amount of time required for growth of lysozyme crystals; 2) X-ray diffraction quality protein crystals can be grown by continuous microwave heating; 3) silver, iron and ITO nanoparticle surfaces can be used to control the growth, size and number of the crystals; and 4) ITO is the most efficient surface for the growth of lysozyme using iCrystal system and the MA-MAEC technique to date.…”

Section: Introductionmentioning

confidence: 99%

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Crystallization of Lysozyme on Metal Surfaces Using a Monomode Microwave System

Mauge-Lewis¹,

Gordon²,

Syed³

et al. 2016

Nano BioMed ENG

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The effect of metal surfaces on the crystallization of lysozyme using the Metal-Assisted and Microwave-Accelerated Evaporative Crystallization (MA-MAEC) technique and a monomode microwave system is described. Our microwave system (is called the iCrystal system hereafter for brevity) is comprised of a 100 W variable power monomode microwave source, a monomode cavity, fiber optic temperature probes and digital cameras. Crystallization of lysozyme (a model protein) was conducted using the iCrystal system on four different types of circular crystallization plates with 21-well sample capacity (i.e., crystallization plates): (i) blank: a continuous surface without a metal, (ii) silver nanoparticle films (SNFs): a discontinuous metal film, (iii) iron nano-columns: a semicontinuous metal film, and (iv) indium tin oxide (ITO): a continuous metal film. Lysozyme crystals grown on all crystallization plates were characterized by X-ray crystallography and found to be X-ray diffraction quality. The use of iron nano-columns afforded for the growth of largest number of lysozyme crystals with a narrow size distribution. ITO-modified crystallization plates were deemed to be best of all the crystallization plates based on the observations that lysozyme crystals were grown at the shortest time (370 ± 36 minutes) with a narrow size distribution up to 460 m in size.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Crystallization of Lysozyme on Metal Surfaces Using a Monomode Microwave System

Mauge-Lewis¹,

Gordon²,

Syed³

et al. 2016

Nano BioMed ENG

View full text Add to dashboard Cite

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“…Only a limited number of methods have been proposed for determining crystal quality prior to diffraction, including analysis of the birefringent properties of protein crystals and low-intensity X-ray diffraction prior to synchrotron X-ray diffraction (Watanabe, 2005;Owen & Garman, 2005). The lack of a reliable bench-top method for rapidly predicting crystal quality adds considerable time and expense to structure-determination efforts, since poorly diffracting low-quality crystals are often only identified as such after crystal harvesting and diffraction analysis by synchrotronradiation X-ray diffraction (Lunde et al, 2005;Vernede et al, 2006;Groves et al, 2007;Garcia-Caballero et al, 2011). During crystal growth, multiple crystals can grow together in nonspecific orientations and can complicate diffraction analysis, often resulting in poor quality of the structural data (Dauter, 2003;Borshchevskiy et al, 2010;Boudjemline et al, 2008;Garcia-Caballero et al, 2011;Yeates & Fam, 1999).…”

Section: Introductionmentioning

confidence: 99%

“…The lack of a reliable bench-top method for rapidly predicting crystal quality adds considerable time and expense to structure-determination efforts, since poorly diffracting low-quality crystals are often only identified as such after crystal harvesting and diffraction analysis by synchrotronradiation X-ray diffraction (Lunde et al, 2005;Vernede et al, 2006;Groves et al, 2007;Garcia-Caballero et al, 2011). During crystal growth, multiple crystals can grow together in nonspecific orientations and can complicate diffraction analysis, often resulting in poor quality of the structural data (Dauter, 2003;Borshchevskiy et al, 2010;Boudjemline et al, 2008;Garcia-Caballero et al, 2011;Yeates & Fam, 1999). Crystalline samples with multiple domains are not always easily identifiable by bright-field imaging, especially for the specific case of twinning.…”

Section: Introductionmentioning

confidence: 99%

Polarization-resolved second-harmonic generation microscopy as a method to visualize protein-crystal domains

DeWalt

Begue

Ronau

et al. 2012

Acta Crystallogr D Biol Cryst

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Polarization-resolved second-harmonic generation (PR-SHG) microscopy is described and applied to identify the presence of multiple crystallographic domains within protein-crystal conglomerates, which was confirmed by synchrotron X-ray diffraction. Principal component analysis (PCA) of PR-SHG images resulted in principal component 2 (PC2) images with areas of contrasting negative and positive values for conglomerated crystals and PC2 images exhibiting uniformly positive or uniformly negative values for single crystals. Qualitative assessment of PC2 images allowed the identification of domains of different internal ordering within protein-crystal samples as well as differentiation between multi-domain conglomerated crystals and single crystals. PR-SHG assessments of crystalline domains were in good agreement with spatially resolved synchrotron X-ray diffraction measurements. These results have implications for improving the productive throughput of protein structure determination through early identification of multi-domain crystals.

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“…as mass availability is no longer a limiting step within the crystallographic project [8,9]. Additionally, the use of synchrotron radiation facilities [10,11] has improved not only the time for data collection (exposing the crystals to less radiation damage), but has also reduced the crystal size (in the scale of microns) needed to obtain X-ray data for structural purposes.…”

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confidence: 99%