Pre-assembled clusters distort crystal nucleation kinetics in supersaturated lysozyme solutions

Parmar, Avanish Singh; Gottschall, Paul E.; Muschol, Martin

doi:10.1016/j.bpc.2007.06.002

Cited by 58 publications

(77 citation statements)

References 52 publications

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“…The effective specific surface energies provided by these authors are quite close (generally, below 1 mJ m −2 ) to the one evaluated here ( Table 2, γ ef ). The activity factor (Table 2, ψ) suggests predominant heterogeneous nucleation, which is in good agreement with the expected natural presence of a variety of impurities in solution samples of commercial lysozyme [19]. The walls of the crystallization solution container have a minor contribution to heterogeneous nucleation, even at a high ratio of wall surface to solution volume [25].…”

Section: Nucleation Parameterssupporting

confidence: 79%

“…More important, however, are the large deviations in t g and the relatively low values of t g , especially at highest lysozyme concentrations. First of all, this could be attributed to the intrinsic impurity content of protein solutions, especially in the considered case, where a variety of aggregates are expected to be present, which are likely to distort nucleation kinetics [19]. The second reason might be in the method of solution preparation.…”

Section: Probability For Nucleationmentioning

confidence: 99%

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Probabilistic approach to lysozyme crystal nucleation kinetics

2015

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Nucleation of lysozyme crystals in quiescent solutions at a regime of progressive nucleation is investigated under an optical microscope at conditions of constant supersaturation. A method based on the stochastic nature of crystal nucleation and using discrete time sampling of small solution volumes for the presence or absence of detectable crystals is developed. It allows probabilities for crystal detection to be experimentally estimated. One hundred single samplings were used for each probability determination for 18 time intervals and six lysozyme concentrations. Fitting of a particular probability function to experimentally obtained data made possible the direct evaluation of stationary rates for lysozyme crystal nucleation, the time for growth of supernuclei to a detectable size and probability distribution of nucleation times. Obtained stationary nucleation rates were then used for the calculation of other nucleation parameters, such as the kinetic nucleation factor, nucleus size, work for nucleus formation and effective specific surface energy of the nucleus. The experimental method itself is simple and adaptable and can be used for crystal nucleation studies of arbitrary soluble substances with known solubility at particular solution conditions.

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Section: Nucleation Parameterssupporting

confidence: 79%

Section: Probability For Nucleationmentioning

confidence: 99%

Probabilistic approach to lysozyme crystal nucleation kinetics

2015

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“…The impurities could be irreversibly aggregated lysozyme molecules. Parmar et al 11 found evidence for non-equilibrium irreversibly formed aggregates of lysozyme, see Fig. 3.…”

Section: Lysozymementioning

confidence: 94%

“…During attempts to crystallise them, solutions of many proteins often separate into dilute and concentrated solution phases. Most of the quantitative studies of this have worked with the small, stable cheap protein lysozyme 11,[92][93][94][95] . Protein crystallisation is of great importance due to the need for protein crystals in order to use X-ray diffraction to determine their structure.…”

Section: Examples Of Crystallisation In the Presence Of A Fluid/fluidmentioning

confidence: 99%

The non-classical nucleation of crystals: microscopic mechanisms and applications to molecular crystals, ice and calcium carbonate

Sear

2012

International Materials Reviews

206

219

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Crystals form via nucleation followed by growth. Often nucleation data is interpreted using the classical theory of nucleation, which is essentially a simple theory for the nucleation of a fluid phase. I characterise this classical theory as making six assumptions; I discuss each assumption in turn. I then review experiments and simulations that find nucleation behaviour that cannot be described by the classical theory. The experiments are on the crystallisation from solution of molecules such as drugs and related molecules, ice and calcium carbonate. The review also covers work on non-classical nucleation in solutions of the protein lysozyme, and work on the fascinating phenomenon of nucleation induced by laser pulses. I hope this review will be of interest to those studying the crystallisation of both molecules and ions from solution. The review aims to advance our understanding of the crucial first step in crystallisation, and to enable researchers studying crystallisation in one system to learn from what others have done in studying analogous phenomena in different systems.

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“…19 Samples were consecutively filtered through 220-nm and 50-nm pore size syringe filters. This concentrated HEWL stock was mixed 1:1 either with a NaCl /25 mM KH 2 PO 4 pH 2 or with a NaCl /20 mM HEPES pH 7 stock solutions, with NaCl concentrations in this salt/buffer stock adjusted to twice their final concentrations.…”

Section: Preparation Of Hewl Growth Solutions and Tht Solutionsmentioning

confidence: 99%

Structural fingerprints and their evolution during oligomeric vs. oligomer-free amyloid fibril growth

Foley

Hill

Miti

et al. 2013

The Journal of Chemical Physics

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Deposits of fibrils formed by disease-specific proteins are the molecular hallmark of such diverse human disorders as Alzheimer's disease, type II diabetes, or rheumatoid arthritis. Amyloid fibril formation by structurally and functionally unrelated proteins exhibits many generic characteristics, most prominently the cross β-sheet structure of their mature fibrils. At the same time, amyloid formation tends to proceed along one of two separate assembly pathways yielding either stiff monomeric filaments or globular oligomers and curvilinear protofibrils. Given the focus on oligomers as major toxic species, the very existence of an oligomer-free assembly pathway is significant. Little is known, though, about the structure of the various intermediates emerging along different pathways and whether the pathways converge towards a common or distinct fibril structures. Using infrared spectroscopy we probed the structural evolution of intermediates and late-stage fibrils formed during in vitro lysozyme amyloid assembly along an oligomeric and oligomer-free pathway. Infrared spectroscopy confirmed that both pathways produced amyloid-specific β-sheet peaks, but at pathwayspecific wavenumbers. We further found that the amyloid-specific dye thioflavin T responded to all intermediates along either pathway. The relative amplitudes of thioflavin T fluorescence responses displayed pathway-specific differences and could be utilized for monitoring the structural evolution of intermediates. Pathway-specific structural features obtained from infrared spectroscopy and Thioflavin T responses were identical for fibrils grown at highly acidic or at physiological pH values and showed no discernible effects of protein hydrolysis. Our results suggest that late-stage fibrils formed along either pathway are amyloidogenic in nature, but have distinguishable structural fingerprints. These pathway-specific fingerprints emerge during the earliest aggregation events and persist throughout the entire cascade of aggregation intermediates formed along each pathway. © 2013 AIP Publishing LLC. [http://dx

show abstract

Pre-assembled clusters distort crystal nucleation kinetics in supersaturated lysozyme solutions

Cited by 58 publications

References 52 publications

Probabilistic approach to lysozyme crystal nucleation kinetics

Probabilistic approach to lysozyme crystal nucleation kinetics

The non-classical nucleation of crystals: microscopic mechanisms and applications to molecular crystals, ice and calcium carbonate

Structural fingerprints and their evolution during oligomeric vs. oligomer-free amyloid fibril growth

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