Lower dimer impurity incorporation may result in higher perfection of HEWL crystals grown in microgravity

Carter, D.C.; Lim, Kian Meng; Ho, Joseph X.; Wright, Brenda S.; Twigg, Pamela D.; Miller, Teresa Y.; Chapman, Jenny; Keeling, Kim M.; Ruble, J. R.; Vekilov, Peter G.; Thomas, B. R.; Rosenberger, Franz; Chernov, A. A.

doi:10.1016/s0022-0248(98)00859-8

Cited by 63 publications

(69 citation statements)

References 38 publications

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“…Another study 10 with CEWL using the Diffusioncontrolled Crystallization Apparatus for Microgravity (DCAM) 37 at ambient temperature aboard the Mir spacestation reported K eff values of 9 on the ground and 2 in microgravity, assuming an initial 1% dimer solution concentration. Given that experimental conditions may influence K eff , it is difficult to make a direct comparison between these two findings.…”

Section: Resultsmentioning

confidence: 99%

“…To date, growth of macromolecular crystals in microgravity has had mixed results, with enhancement in some studies [2][3][4][5] and no positive or even detrimental effects in others. [6][7][8] CEWL, a frequent flyer in microgravity crystallization programs, reflects this variation with reports of decreased mosaicity, 4,9 increased diffraction resolution, 10 marginal improvement, 11 and no effects. [7][8] In biological crystal growth, macromolecule purity is frequently given as a major parameter governing success in crystallization.…”

Section: Introductionmentioning

confidence: 99%

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Investigating the Effect of Impurities on Macromolecule Crystal Growth in Microgravity

Snell

Judge

Crawford

et al. 2001

Crystal Growth & Design

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Chicken egg-white lysozyme (CEWL) crystals were grown in microgravity and on the ground in the presence of various amounts of a naturally occurring lysozyme dimer impurity. No significant favorable differences in impurity incorporation between microgravity and ground crystal samples were observed. At low impurity concentration the microgravity crystals preferentially incorporated the dimer. The presence of the dimer in the crystallization solutions in microgravity reduced crystal size, increased mosaicity, and reduced the signal-to-noise ratio of the X-ray data. Microgravity samples proved more sensitive to impurity. Accurate indexing of the reflections proved critical to the X-ray analysis. The largest crystals with the best X-ray diffraction properties were grown from pure solution in microgravity.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Investigating the Effect of Impurities on Macromolecule Crystal Growth in Microgravity

Snell

Judge

Crawford

et al. 2001

Crystal Growth & Design

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“…[17][18][19][20][21][22][23][24] Generally, impurities are considered to exhibit their effects on the growth process after they adsorb on a crystal surface. Thus, to fully comprehend the mechanisms of impurity effects, one has to observe in situ both (1) dynamics of elementary steps and (2) adsorption of impure molecules on a crystal surface, at a molecular level.…”

Section: Introductionmentioning

confidence: 99%

Direct Observation of Adsorption Sites of Protein Impurities and Their Effects on Step Advancement of Protein Crystals

Sazaki

戴国亮

et al. 2009

Crystal Growth & Design

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ABSTRACT:We measured noninvasively step velocities of elementary two-dimensional (2D) islands on {110} faces of tetragonal lysozyme crystals, under various supersaturations, by laser confocal microscopy combined with differential interference contrast microscopy. We studied the correlation between the effects of protein impurities on the growth of elementary steps and their adsorption sites on a crystal surface, using three kinds of proteins: fluorescent-labeled lysozyme (F-lysozyme), covalently bonded dimers of lysozyme (dimer), and a 18 kDa polypeptide (18 kDa). These three protein impurities suppressed the advancement of the steps. However, they exhibited different supersaturation dependencies of the suppression of the step velocities. To clarify the cause of this difference, we observed in situ the adsorption sites of individual molecules of F-lysozyme and fluorescent-labeled dimer (F-dimer) on the crystal surface by single-molecule visualization. We found that F-lysozyme adsorbed preferentially on steps (i.e., kinks), whereas F-dimer adsorbed randomly on terraces. Taking into account the different adsorption sites of F-lysozyme and F-dimer, we could successfully explain the different effects of the impurities on the step velocities. These observations strongly suggest that 18 kDa also adsorbs randomly on terraces. Seikagaku lysozyme exhibited a complex effect that could not alone be explained by the two major impurities (dimer and 18 kDa) present in Seikagaku lysozyme, indicating that trace amounts of other impurities significantly affect the step advancement.

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“…These impurities could be incorporated into protein crystals to form lattice defects. In the last decades, therefore, numerous studies about impurity effects on protein crystal growth have been carried out [2][3][4][5][6]. Vekilov et al [7] reported the impurity effect on the growth step kinetics.…”

Section: Introductionmentioning

confidence: 99%

Impurity effect on defect formation of protein crystals

Hondoh

Nakada

2005

Journal of Crystal Growth

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Lower dimer impurity incorporation may result in higher perfection of HEWL crystals grown in microgravity

Cited by 63 publications

References 38 publications

Investigating the Effect of Impurities on Macromolecule Crystal Growth in Microgravity

Investigating the Effect of Impurities on Macromolecule Crystal Growth in Microgravity

Direct Observation of Adsorption Sites of Protein Impurities and Their Effects on Step Advancement of Protein Crystals

Impurity effect on defect formation of protein crystals

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