Association-Dissociation Properties of Lysozyme<sup>*</sup>

Bruzzesi, M. Rosaria; Chiancone, Emilia; Antonini, Eraldo

doi:10.1021/bi00885a016

Cited by 108 publications

(48 citation statements)

References 9 publications

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“…Lysozyme is known to undergo reversible selfassociation at pH . 5 and in highly concentrated solution (Chiancone et al 1965;Gottschalk & Halle 2003). In any case, we observed that the measurement performed after the pressure release reproduces the same diffusion coefficient (in general, the same S(Q, v) of the first measurement), suggesting a complete reversibility of the phenomenon.…”

Section: Dynamics: Quasi-elastic Neutron Scatteringsupporting

confidence: 63%

Combining structure and dynamics: non-denaturing high-pressure effect on lysozyme in solution

Ortore

Spinozzi

Mariani

et al. 2009

J. R. Soc. Interface.

130

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Small-angle X-ray scattering (SAXS) and elastic and quasi-elastic neutron scattering techniques were used to investigate the high-pressure-induced changes on interactions, the low-resolution structure and the dynamics of lysozyme in solution. SAXS data, analysed using a global-fit procedure based on a new approach for hydrated protein form factor description, indicate that lysozyme completely maintains its globular structure up to 1500 bar, but significant modifications in the protein -protein interaction potential occur at approximately 600-1000 bar. Moreover, the mass density of the protein hydration water shows a clear discontinuity within this pressure range. Neutron scattering experiments indicate that the global and the local lysozyme dynamics change at a similar threshold pressure. A clear evolution of the internal protein dynamics from diffusing to more localized motions has also been probed. Protein structure and dynamics results have then been discussed in the context of protein -water interface and hydration water dynamics. According to SAXS results, the new configuration of water in the first hydration layer induced by pressure is suggested to be at the origin of the observed local mobility changes.

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Section: Dynamics: Quasi-elastic Neutron Scatteringsupporting

confidence: 63%

Combining structure and dynamics: non-denaturing high-pressure effect on lysozyme in solution

Ortore

Spinozzi

Mariani

et al. 2009

J. R. Soc. Interface.

130

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show abstract

“…Multiple peaks and merging of peaks due to aggregation have also been observed in electrophoresis of biological molecules (Cann, and Toral, 1989). Some traditional techniques for studying aggregation behavior are sedimentation and low-angle laser light scattering (LALLS) (Bruzzesi et al, 1965;Stuting et al, 1989;Mhatre et al, 1990). Kihara and Tsuruta (1988) have characterized both aggregation kinetics and conformational changes by stopped flow X-ray scattering.…”

Section: General Characteristics Of Aggregation In Separationsmentioning

confidence: 99%

Effects of protein aggregation in isocratic nonlinear chromatography

et al. 1991

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“…Chromatography of lysozyme on a column eyuilibrated and developed with 1 mM N-acetylglucosamine showed a decrease of 99.l0/, in the free form in favour of the adsorbed form (Table 1), despite the accompanying increase in ionic strength to 10-3 which would normally favour the free form. It appears that the adsorbed form is the monomer, and the free form is polymeric since the inhibition of associa- A monomer-dimer-trimer system has been proposed [6]; it has also been reported that a t I = 0.3 and higher lysozyme concentration, higher species appear [3]. The linear increase in equilibrium ratio values with the rise of concentration (Fig.2) suggested that the association should be interpreted in terms of more than one linked equilibria.…”

Section: Resultsmentioning

confidence: 99%

Conformation of Lysozyme in Aqueous Solution

Hampe

1972

European Journal of Biochemistry

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The ionic strength, concentration, and temperature effects on the association-dissociation properties of lysozyme in aqueous solution have been studied by gel chromatography. The following results were obtained:1. Increasing ionic strength leads to a n increase in inter-conversion rate between the monomeric and associated forms. Between distilled water and 0.01 ionic strength, lysozyme shows a low velocity of interconversion, whereas between 0.01 and 0.2 rates are observed which are rapid as compared with the column fractionation time.2. Analysis of concentration dependence indicates the formation of more highly associated forms than t,he dimer. The data can be fitted within experimental error by an indefinite association with the same equilibrium constant for the addition of each successive monomer.3. The equilibrium ratio for the slow polymerization process measured a t temperatures between 15 ' and 40 "C yields the apparent thermodynamic quantities, AGO, AHo, and AS0 for the association reaction. This data is consistent with a hydrophobic interaction between lysozyme monomers and Sephadex gel matrix. It is suggested that the adsorption involves the tryptophan residues of the active centre. The enthalpy of the polymerization process is interpreted in terms of a conformational change of the same order of magnitude as on the bonding of N-acetylglucosamine to the enzyme.Lysozyme has long been recognized as a protein that self-associates in aqueous solution [l--61. During chromatography on a Sephadex G-50 column equilibrated with distilled water, part of the lysozyme is eluted, and part, which may subsequently be eluted with 0. Definitions. req, equilibrium ratio of adsorption; A,,,, unit, amount of material which when dissolved in 1 ml gives an absorbance of 1 in a I-cm pathlength cell.Enzyme. Lysozyme (EC 3.2.1.17).[3]. Application of the multinomial theory suggests for lysozyme, a t 4 "C and pH 7.2, an ideal monomerdimer-trimer associating system [6].Experiments have been done in order to elucidate more clearly the effect of ionic strength, protein concentration and temperature on the aggregation status of lysozyme in aqueous solution. MATERIALS AND METHODSHen egg-white lysozyme from Nutritional Biochemicals (lot No. 6573, three-times crystallized, from Sigma, 22500 U/mg L-100 B-92) and four-times crystallized lysozyme, prepared as described previously [12], were used. Most experiments were carried out with lysozyme from Nutritional Biochemicals, a few with Sigma and that prepared in the laboratory, but, no significant differences were observed.Chromatography was performed using Sephadex G-15, 6-25, and G-50 fine from Pharmacia (Uppsala, Sweden) and Bio-Gel P-6 from Bio-Rad in a jacketed column (1 x 30 cm) which was maintained a t constant temperature by circulation of water. The gel column was equilibrated and developed with the same solvent. The solvent used were: distilled water, sodium chloride solutions of different ionic strength,

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Association-Dissociation Properties of Lysozyme^*

Cited by 108 publications

References 9 publications

Combining structure and dynamics: non-denaturing high-pressure effect on lysozyme in solution

Combining structure and dynamics: non-denaturing high-pressure effect on lysozyme in solution

Effects of protein aggregation in isocratic nonlinear chromatography

Conformation of Lysozyme in Aqueous Solution

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Association-Dissociation Properties of Lysozyme*

Cited by 108 publications

References 9 publications

Combining structure and dynamics: non-denaturing high-pressure effect on lysozyme in solution

Combining structure and dynamics: non-denaturing high-pressure effect on lysozyme in solution

Effects of protein aggregation in isocratic nonlinear chromatography

Conformation of Lysozyme in Aqueous Solution

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Association-Dissociation Properties of Lysozyme^*