Direct Crystallization of Lysozyme From Egg White and Some Crystalline Salts of Lysozyme

Alderton, Gordon; Fevold, H. L.

doi:10.1016/s0021-9258(18)43040-2

Cited by 253 publications

(44 citation statements)

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“…Our results also show a substantial effect of pH on crystal numbers with a change of two orders of magnitude over the pH range 4.0 -5.2. Alderton and Fevold (1946), with initial lysozyme concentrations of 40 mg/ml in unbuffered 5% NaCl solutions, may be referring to this when they describe the crystallization to be copious at pH 3.5, moderate at pH 4.8, and very slight at pH 5.8. This effect of pH on crystal numbers appears to be independent of solution pH history.…”

Section: Discussionmentioning

confidence: 99%

“…Ries- Kautt and Ducruix (1992) made the general observation that lysozyme crystals appeared faster and in larger numbers with increasing supersaturation. Alderton and Fevold (1946) reported that in initial lysozyme concentrations of 40 mg/ml, in 5% NaCl solutions, crystallization was copious at pH 3.5, moderate at pH 4.8, and very slight at pH 5.8, indicating a strong effect of pH. Elgersma et al (1992) (their figure 4) also report an effect of pH on lysozyme crystal numbers, as large numbers of crystals can be seen at pH 4.0 (ln(c/s) ϭ 1.2), whereas fewer crystals are present at pH 6.0 (ln(c/s) ϭ 1.6).…”

Section: Introductionmentioning

confidence: 99%

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The Effect of Temperature and Solution pH on the Nucleation of Tetragonal Lysozyme Crystals

Judge

Jacobs

Frazier

et al. 1999

Biophysical Journal

110

113

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Part of the challenge of macromolecular crystal growth for structure determination is obtaining crystals with a volume suitable for x-ray analysis. In this respect an understanding of the effect of solution conditions on macromolecule nucleation rates is advantageous. This study investigated the effects of supersaturation, temperature, and pH on the nucleation rate of tetragonal lysozyme crystals. Batch crystallization plates were prepared at given solution concentrations and incubated at set temperatures over 1 week. The number of crystals per well with their size and axial ratios were recorded and correlated with solution conditions. Crystal numbers were found to increase with increasing supersaturation and temperature. The most significant variable, however, was pH; crystal numbers changed by two orders of magnitude over the pH range 4.0-5.2. Crystal size also varied with solution conditions, with the largest crystals obtained at pH 5.2. Having optimized the crystallization conditions, we prepared a batch of crystals under the same initial conditions, and 50 of these crystals were analyzed by x-ray diffraction techniques. The results indicate that even under the same crystallization conditions, a marked variation in crystal properties exists.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The Effect of Temperature and Solution pH on the Nucleation of Tetragonal Lysozyme Crystals

Judge

Jacobs

Frazier

et al. 1999

Biophysical Journal

110

113

View full text Add to dashboard Cite

show abstract

“…Crystals of hen egg-white lysozyme, grown a t pH 4*7 (Alderton & Fevold 1946), are tetragonal with a = b = 79*1 A, c = 37*9 A, s Ballantyre & Galvin 1948; Blake, Fenn, North, Phillips & Poljak 1962). Each of the eight asymmetric units in the cell comprises a single lysozyme molecule, molecular weight about 14600, together with 1 M sodium chloride solution which constitutes some 33*5 % of the weight of the crystal (Steinrauf 1959).…”

mentioning

confidence: 99%

On the conformation of the hen egg-white lysozyme molecule

Blake

Mair

North

et al. 1967

Proc. R. Soc. Lond. B.

379

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Crystals of hen egg-white lysozyme, grown at pH 4.7 (Alderton & Fevold 1946), are tetragonal with a = b = 79.1 Å, c = 37.9 Å, space group P 4 3 2 1 2 (Palmer, Ballantyre & Galvin 1948; Blake, Fenn, North, Phillips & Poljak 1962). Each of the eight asymmetric units in the cell comprises a single lysozyme molecule, molecular weight about 14 600, together with 1 M sodium chloride solution which constitutes some 33.5% of the weight of the crystal (Steinrauf 1959). The structure of these crystals has been determined by X-ray analysis by the method of multiple isomorphous replacement developed in the studies of haemoglobin (Green, Ingram & Perutz 1954; Blow 1958; Perutz, Rossmann, Cullis, Muirhead, Will & North 1960) and myoglobin (Kendrew, Dickerson, Strandberg, Hart, Davies, Phillips & Shore 1960). Anomalous scattering data were used in conjunction with the isomorphous replacement intensity differences (North 1965) to form a joint probability distribution for the phase of each reflexion. The position of the centroid of each probability distribution gave a phase angle and weighting factor for each reflexion from which the electron density map with minimum r.m.s. error was calculated (Blow & Crick 1959). A large number of different heavy atom derivatives were studied (Poljak 1963; Blake, Koenig, Mair, North, Phillips & Sarma 1965) and three proved satisfactory for calculating an electron density map at 2 Å resolution. They contained respectively ortho -mercuri hydroxytoluene para -sulphonic acid, UO 2 F 5 3- and an ion derived from UO 2 (NO 3 ) 2 , probably UO 2 (OH) n (n-2)-

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“…Lysozyme in the tetragonal form [17] can be reproducibly grown at a pH between 4.0 and 7.5, canavalin between about pH 5.0 and 7.5 [14][15][16], and thaumatin [18] between pH 6.0 and 8.0. Although crystals of trypsin, inhibited with benzamidine [19], are usually grown at pH 8.5, they can be transferred to mother liquors at pH 6.0 and even below, without damage.…”

Section: Methodsmentioning

confidence: 99%

pH and Redox Induced Color Changes in Protein Crystals Suffused with Dyes

McPherson¹

2019

Crystals

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Protein crystals, otherwise usually colorless, can be stained a variety of hues by saturating them with dyes, by diffusion from the mother liquor or co-crystallization. The colors assumed by dyes are a function of chemical factors, particularly pH and redox potential. Protein crystals saturated with a pH sensitive dye, initially at one pH, can be exposed to the mother liquor at a second pH and the crystal will change color over time as H3O+ ions diffuse through the crystal. This allows diffusion rates of H3O+ through the crystal to be measured. Diffusion fronts are often clearly delineated. Similar experiments can be carried out with redox sensitive dyes by adding reductants, such as ascorbic acid or dithionite, or oxidants such as H2O2, to the crystal’s mother liquor. Presented here are a number of experiments using pH or redox sensitive dye-saturated protein crystals, and some experiments using double dye, sequential redox–pH changes.

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Direct Crystallization of Lysozyme From Egg White and Some Crystalline Salts of Lysozyme

Cited by 253 publications

References 2 publications

The Effect of Temperature and Solution pH on the Nucleation of Tetragonal Lysozyme Crystals

The Effect of Temperature and Solution pH on the Nucleation of Tetragonal Lysozyme Crystals

On the conformation of the hen egg-white lysozyme molecule

pH and Redox Induced Color Changes in Protein Crystals Suffused with Dyes

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