Effects of purification on the crystallization of lysozyme

Ewing, F.; Forsythe, Elizabeth L.; Woerd, Mark van der; Pusey, Marc L.

doi:10.1016/0022-0248(95)00933-7

Cited by 38 publications

(35 citation statements)

References 31 publications

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“…Table 1 crystallographic data obtained. The unit-cell dimensions for tetragonal lysozyme crystals prepared from ammonium sulfate at pH 4.4, a = b = 78.71, c = 38.6 A, were slightly outside the ranges previously reported for this morphology, a = b = 78.83-79.23, c --37.89-38.5 A (Palmer, Ballantyne & Galvin, 1948;Steinrauf, 1959;Joll~s & Berthou, 1972;Berthou & Joll~s, 1974;Ri~s-Kautt & Ducruix, 1989;Ri~s-Kautt et al, 1994;Ewing, Forsythe, van der Woerd & Pusey, 1996 (Alderton & Fevold, 1946;Crick, 1953;Steinrauf, 1959;Haas, 1967;Hogle et al, 1981;Rirs-Kautt & Ducruix, 1989;Guilloteau et al, 1992). Steinrauf (1959) obtained fragile monoclinic plates using 10% (0.7M) sodium sulfate, but in the presence of 1.0M acetate buffer.…”

Section: Resultsmentioning

confidence: 54%

Crystallization of chicken egg-white lysozyme from ammonium sulfate

Forsythe¹,

Snell²,

Pusey³

1997

Acta Crystallogr D Biol Crystallogr

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Chicken egg-white lysozyme was crystallized from ammonium sulfate over the pH range 4.0-7.8, with protein concentrations from 100 to 150 mgml 1. Crystals were obtained by vapordiffusion or batch-crystallization methods. The protein crystallized in two morphologies with an apparent morphology dependence on temperature and protein concentration. In general, tetragonal crystals could be grown by lowering the protein concentration or temperature. Increasing the temperature or protein concentration resulted in the growth of orthorhombic crystals. Representative crystals of each morphology were selected for X-ray analysis. The tetragonal crystals belonged to the P432~2 space group with crystals grown atoPH 4.4 having unit-cell dimensions of a = b= 78.71, c= 38.6 A and diffracting to beyond 2.0 A. The orthorhombic crystals, grown at pH 4.8, were of space group P21212 and had unit-cell dimensions of a = 30.51, b = 56.51 and c = 73.62 ,~.

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Section: Resultsmentioning

confidence: 54%

Crystallization of chicken egg-white lysozyme from ammonium sulfate

Forsythe¹,

Snell²,

Pusey³

1997

Acta Crystallogr D Biol Crystallogr

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“…The powdery solids that formed frequently at 5% and 6% NaCl had a 'sea urchin' morphology at lower magnification, but clearly had faceted edges when viewed at higher magnification, 400 Â , as shown in. material appeared as clumps with 'sea-urchin' morphology reported by others in crystallizing lysozyme [14][15][16]. However, under high magnification, the powdery material was seen to consist of needles with regular structure and sharp, faceted edges, approximately 5-50 mm in the longest dimension.…”

Section: Resultsmentioning

confidence: 73%

Dependence of nucleation kinetics and crystal morphology of a model protein system on ionic strength

Bhamidi

Skrzypczak‐Jankun

Schall

2001

Journal of Crystal Growth

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Nucleation rate data for hen egg-white lysozyme crystallization were obtained using a particle counter. Tetragonal lysozyme crystals were expected to form at the temperature and solution conditions of these experiments: 48C, pH 4.5 with 0.1 M sodium acetate buffer and 2-6% NaCl (w/v). The rates varied as expected, as smooth monotonic functions of supersaturation at 2%, 3% and 6% NaCl. However, at 5% NaCl, a great deal of scatter in the data was observed. At 2% and 3% NaCl, all the batches contained crystals with tetragonal morphology. At 6% NaCl, almost all of the vials contained the white powder with few or no tetragonal crystals. At 5% NaCl concentration, a mixture of tetragonal crystals and powder formed in varying proportions in all the vials as observed by visual inspection. The powdery material was examined using optical microscopy and was seen to consist of needles with regular structure and sharp, faceted edges. Powder diffraction data from these needles was inconsistent with experimental powder diffraction data from tetragonal lysozyme crystals. It is possible that at high salt and protein concentrations liquid-liquid separation occurred and yielded a crystal polymorph. #

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“…Multiple investigators have reported contamination of lyophilized lysozyme stocks by low molecular weight (b 100 kD) protein impurities [16,47,51]. We set out to determine whether the pre-assembled clusters found in lyophilized lysozyme stocks were indeed protein clusters, and which proteins they were composed of.…”

Section: Analysis Of Cluster Compositionmentioning

confidence: 99%

“…Protein impurities have also been implicated in changes to growth rates on the 101 facet of tetragonal lysozyme crystals [12,34,50], the density of optical defects in lysozyme crystals [6] and limitations of X-ray diffraction resolution [7,33,43]. Even in the absence of contaminating impurities, structural micro-heterogeneities of lysozyme monomers have been linked to altered crystal habit and crystal quality [16]. These report, however, implicitly assumed that the contaminants were foreign proteins, or small oligomers of lysozyme.…”

Section: Introductionmentioning

confidence: 95%