Crystal growth studies of lysozyme as a model for protein crystallization

Durbin, Stephen D.; Fehér, G.

doi:10.1016/0022-0248(86)90175-2

Cited by 246 publications

(202 citation statements)

References 24 publications

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“…The nucleated crystal is rough, due to high local supersaturation encountered during nucleation and growth. Growth rate (between 600 and 10800s) is 15μm/h in agreement with the growth rate obtained by Durbin et al [63] at high supersaturation for lysozyme. Compared to optical determination of the induction time, the measures of the current during the experiment are clearly more sensitive (Fig.…”

Section: Electrical Field and Confinement By Gelsupporting

confidence: 91%

Addressing the Stochasticity of Nucleation: Practical Approaches

Candoni

Hammadi

Grossier

et al. 2015

Advances in Organic Crystal Chemistry

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Section: Electrical Field and Confinement By Gelsupporting

confidence: 91%

Addressing the Stochasticity of Nucleation: Practical Approaches

Candoni

Hammadi

Grossier

et al. 2015

Advances in Organic Crystal Chemistry

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“…1). The morphologies of the crystals were similar to those of corresponding macroscopic crystals that form under similar conditions (14,15). For additional identification, we analyzed the electron diffraction patterns generated by these crystalline phases ( Fig.…”

Section: Significancementioning

confidence: 82%

Two types of amorphous protein particles facilitate crystal nucleation

Yamazaki

Kimura

Vekilov

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

163

165

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Nucleation, the primary step in crystallization, dictates the number of crystals, the distribution of their sizes, the polymorph selection, and other crucial properties of the crystal population. We used timeresolved liquid-cell transmission electron microscopy (TEM) to perform an in situ examination of the nucleation of lysozyme crystals. Our TEM images revealed that mesoscopic clusters, which are similar to those previously assumed to consist of a dense liquid and serve as nucleation precursors, are actually amorphous solid particles (ASPs) and act only as heterogeneous nucleation sites. Crystalline phases never form inside them. We demonstrate that a crystal appears within a noncrystalline particle assembling lysozyme on an ASP or a container wall, highlighting the role of heterogeneous nucleation. These findings represent a significant departure from the existing formulation of the two-step nucleation mechanism while reaffirming the role of noncrystalline particles. The insights gained may have significant implications in areas that rely on the production of protein crystals, such as structural biology, pharmacy, and biophysics, and for the fundamental understanding of crystallization mechanisms.nucleation | protein | lysozyme | transmission electron microscopy | in situ observation C rystallization can be divided into two processes: nucleation and crystal growth. The crystal growth process has been well examined for a long time, yet the nucleation process is not understood; for example, the nucleation rate of crystals provides a textbook example of order-of-magnitude discrepancies between theoretical predictions and experimental results. Recent proposals have attributed these discrepancies to a nonclassical nucleation pathway, along which a structured crystalline embryo forms within a highly concentrated disordered precursor (1). This mechanism was first proposed for protein crystals (2, 3). Direct observations have demonstrated its applicability to organic (4), inorganic (5, 6), and colloidal (7) crystals. In proteins, clusters of protein molecules have been suggested as precursors; these clusters have mesoscopic sizes from several tens to several hundreds of nanometers and are considered to behave like liquids. It has also been suggested that the precursor is thermodynamically stable with respect to the mother liquid phase but is metastable or unstable with respect to the crystalline phase. The latter nature of the precursor differs from the stable macroscopically dense liquid formed as a result of the liquid-liquid phase separation (8). Such protein-rich mesoscopic clusters have been observed for many proteins, primarily using optical techniques, and have been tentatively identified as precursors for crystal nucleation (9-12). Several important questions concerning this mechanism remain unanswered. First, are the observed mesoscopic clusters actually liquid-like or solid-amorphous? Second, do they play an active role in crystal nucleation? In addition, finally, do the clusters serve as classical heteroge...

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“…at lower supersaturations, when compared with the smoother (110) face (Durbin & Feher, 1986;Monaco & Rosenberger, 1993;Forsythe & Pusey, 1994). Perhaps more significantly, the smoothness of the (110) face is indicated by its slower dissolution rates during etching of the crystal (Monaco & Rosenberger, 1993).…”

Section: Molecular Packing On Edges and Verticesmentioning

confidence: 99%

Growth Mechanism and Morphology of Tetragonal Lysozyme Crystals

Nadarajah¹,

Pusey²

1996

Acta Cryst D

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The tetragonal form of hen egg-white lysozyme is the most investigated protein crystal for growth studies, but the relationship between its surface morphology and internal structure is still not well understood. One method of determining this relationship for inorganic crystals is by employing the periodic bond chain (PBC) theory of Hartman & Perdok [Hartman & Perdok (1955).Acta Co'st. 8,[49][50][51][52][521][522][523][524][525][526][527][528][529]. However, complexities resulting from the packing arrangements and the number of intermolecular bonds in protein crystals have resulted in the use of only simplified versions of this theory so far. In this study a more complete PBC analysis of tetragonal lysozyme crystals was carried out, coupled with an approach incorporating the molecular orientations of the crystal structure. The analysis revealed the existence of a helical tetramer building block of the entire crystal structure, centered around the 43 crystallographic axes, resulting in doublelayered slices and PBC's throughout. The analysis also indicated that the crystallizing units for the faces are at least as large as this tetramer, with the experimental evidence suggesting that it is a tetramer unit for the { 101 } faces and an octamer unit for the {110} faces. The { 110} faces were shown to be molecularly smooth F faces, while the { 101 } to be essentially rough S faces. The predicted morphology and growth mechanisms were found to explain numerous experimental observations from electron and atomic force microscopy, etching studies, lysozyme aggregation studies and measurements of growth kinetics.

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Crystal growth studies of lysozyme as a model for protein crystallization

Cited by 246 publications

References 24 publications

Addressing the Stochasticity of Nucleation: Practical Approaches

Addressing the Stochasticity of Nucleation: Practical Approaches

Two types of amorphous protein particles facilitate crystal nucleation

Growth Mechanism and Morphology of Tetragonal Lysozyme Crystals

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