Structural effects of monovalent anions on polymorphic lysozyme crystals

Vaney, M.C.; Broutin, Isabelle; Retailleau, Pascal; Douangamath, A.; Lafont, Sylvaine; Hamiaux, Cyril; Prangé, Thierry; Ducruix, A.; Riès-Kautt, Madeleine

doi:10.1107/s0907444901004504

Cited by 97 publications

(120 citation statements)

References 35 publications

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“…Detection of anions by X-ray crystallography confirms the penetration of the first hydration layer by counter-ions (Vaney et al, 2001). It remains to fully understand the microscopic interactions that account for the macroscopic properties.…”

Section: Resultsmentioning

confidence: 94%

Importance of the nature of anions in lysozyme crystallisation correlated with protein net charge variation

Retailleau¹,

Ducruix²,

Riès-Kautt³

2002

Acta Crystallogr D Biol Cryst

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Hofmeister anion series are studied to examine the coupled influence of pH and ionic strength on the solubility of previously desalted lysozyme. Solubility curves are measured at pH 4.3 and 8.3 and 18°C for nitrate, para-toluenesulfonate, citrate, sulphate, phosphate, and acetate. Extreme low ionic strength is explored, confirming the decrease of lysozyme solubility while increasing the protein net charge and the ionic strength. The classification of specific salt effects takes into account the valence of the anions with respect to the protein net charge.

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Section: Resultsmentioning

confidence: 94%

Importance of the nature of anions in lysozyme crystallisation correlated with protein net charge variation

Retailleau¹,

Ducruix²,

Riès-Kautt³

2002

Acta Crystallogr D Biol Cryst

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“…It should also be noted that bigger anions also have fewer water molecules in their hydration shells (45). X-ray crystallography has revealed the presence of associated anions in lysozyme crystals when they are grown in the presence of the corresponding salt solution (46,47). The anions are not randomly distributed on protein surface, but instead prefer to interact with specific hydrophilic residues, especially those which are positive charged such as Arg and Lys (46,47).…”

Section: Resultsmentioning

confidence: 99%

“…X-ray crystallography has revealed the presence of associated anions in lysozyme crystals when they are grown in the presence of the corresponding salt solution (46,47). The anions are not randomly distributed on protein surface, but instead prefer to interact with specific hydrophilic residues, especially those which are positive charged such as Arg and Lys (46,47). The distance between individual anions and the protein binding sites in the crystal structures indicate that intervening water is not present, although water molecules may be directly involved in anion-lysozyme interactions in solution.…”

Section: Resultsmentioning

confidence: 99%

The inverse and direct Hofmeister series for lysozyme

Zhang

Cremer

2009

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

391

542

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Anion effects on the cloud-point temperature for the liquid؊liquid phase transition of lysozyme were investigated by temperature gradient microfluidics under a dark field microscope. It was found that protein aggregation in salt solutions followed 2 distinct Hofmeister series depending on salt concentration. Namely, under low salt conditions the association of anions with the positively charged lysozyme surface dominated the process and the phase transition temperature followed an inverse Hofmeister series. This inverse series could be directly correlated to the size and hydration thermodynamics of the anions. At higher salt concentrations, the liquid-liquid phase transition displayed a direct Hofmeister series that correlated with the polarizability of the anions. A simple model was derived to take both charge screening and surface tension effects into account at the protein/water interface. Fitting the thermodynamic data to this model equation demonstrated its validity in both the high and low salt regimes. These results suggest that in general positively charged macromolecular systems should show inverse Hofmeister behavior only at relatively low salt concentrations, but revert to a direct Hofmeister series as the salt concentration is increased.liquid-liquid phase transition ͉ protein aggregation P rotein-protein interactions can lead to aggregation. Such intermolecular contacts underlie the mechanistic basis for a variety of diseases (1-4) and are key to protein crystallization (5-7). An effective way to determine the strength of biomacromolecular interactions is to study the temperature-induced phase separation that occurs in concentrated protein solutions (8). When a concentrated protein solution is cooled below its cloud-point temperature, the system can separate into 2 coexisting liquid phases: 1 rich in protein and 1 poor. As coacervate droplets of the protein-rich phase grow, the solution turns cloudy (white) as a result of the scattering of visible light. Upon standing, the solution may completely separate by gravity into a protein-rich phase and a nearly pure aqueous phase above it. The temperature at which the initial cloud point occurs provides a simple physical measurement of the forces acting among biomacromolecules. Specifically, the higher the temperature at which the initial cloud point occurs, the stronger the putative attractive forces between the protein molecules should be.Dissolved salts in aqueous solutions have a strong influence on protein-protein interactions and the subsequent aggregated states which are formed. In fact, cloud-point temperatures typically follow a specific ion order according to the Hofmeister series (5,6,(9)(10)(11)(12)(13)(14). The relative effectiveness of anions to induce protein aggregation typically follows 1 of 2 trends depending on the pH of the solution (15, 16). Above a protein's isoelectric point, the macromolecule bears a net negative charge and a direct Hofmeister series is normally observed. In this case, chaotropes such as I Ϫ , ClO 4 Ϫ , and SCN Ϫ he...

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“…The number of nitrate ions located in the six molecules varies between ®ve and eight. In a recent study, Vaney et al (2001) have identi®ed seven sites for nitrate ions in monoclinic lysozyme, some often occupied by more than one nitrate ion. All the nitrate ions identi®ed in the present study occupy one or the other of the seven identi®ed sites.…”

Section: Bound Nitrate Ionsmentioning

confidence: 99%

Effect of stabilizing additives on the structure and hydration of proteins: a study involving monoclinic lysozyme

Saraswathi

Sankaranarayanan

Vijayan

2002

Acta Crystallogr D Biol Cryst

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In pursuance of a long-range programme on the hydration, mobility and action of proteins, the structural basis of the stabilizing effect of sugars and polyols is being investigated. With two crystallographically independent molecules with slightly different packing environments in the crystal, monoclinic lysozyme constitutes an ideal system for exploring the problem. The differences in the structure and hydration of the two molecules provide a framework for examining the changes caused by stabilizing additives. Monoclinic crystals were grown under native conditions and also in the presence of 10% sucrose, 15% trehalose, 10% trehalose, 10% sorbitol and 5% glycerol. The crystal structures were re®ned at resolutions ranging from 1.8 to 2.1 A Ê . The average B values, and hence the mobility of the structure, are lower in the presence of additives than in the native crystals. However, a comparison of the structures indicates that the effect of the additives on the structure and the hydration shell around the protein molecule is considerably less than that caused by differences in packing. It is also less than that caused by the replacement of NaNO 3 by NaCl as the precipitant in the crystallization experiments. This result is not in conformity with the commonly held belief that additives exert their stabilizing effect through the reorganization of the hydration shell, at least as far as the ordered water molecules are concerned. PDB References: lysozyme, native monoclinic (I), 1lj3, r1lj3sf; native monoclinic (II), 1lj4, r1lj4sf; with 10% sucrose (I), 1lje, r1ljesf; with 10% sucrose (II), 1ljf, r1ljfsf; with 5% glycerol (I), 1ljg, r1ljgsf; with 5% glycerol (II), 1ljh, r1ljhsf; with 10% sorbitol, 1lji, r1ljisf; with 10% trehalose, 1ljj, r1ljjsf; with 15% trehalose, 1ljk, r1ljksf.

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Structural effects of monovalent anions on polymorphic lysozyme crystals

Cited by 97 publications

References 35 publications

Importance of the nature of anions in lysozyme crystallisation correlated with protein net charge variation

Importance of the nature of anions in lysozyme crystallisation correlated with protein net charge variation

The inverse and direct Hofmeister series for lysozyme

Effect of stabilizing additives on the structure and hydration of proteins: a study involving monoclinic lysozyme

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