A novel approach for the crystallization of soluble proteins using non-ionic surfactants

Mustafa, Aziz O.; Derrick, Jeremy P.; Tiddy, G. J. T.; Ford, Robert C.

doi:10.1107/s0907444997009074

Cited by 6 publications

(7 citation statements)

References 16 publications

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“…One particular class of such additives is surfactants, which are known to bind to proteins in solution, , and in larger quantities, to cause their denaturation . Small amounts of nonionic surfactants may improve the quality of crystals formed from soluble proteins, but under other conditions can cause precipitation by modification of the protein−protein interactions. ,,, We have found that the presence of small quantities of anionic surfactants can drastically modify the morphologies of protein crystalline phases . Both the continuum properties and the molecular mechanisms of surfactant action during protein crystallization are, however, poorly characterized.…”

Section: Introductionmentioning

confidence: 96%

Surfactant Diffusion into Lysozyme Crystal Matrices Investigated by Quantitative Fluorescence Microscopy

2000

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Small quantities of ionic surfactants modify the crystallization of lysozyme and lead to the formation of phases with different morphologies. By using fluorescent probes, we demonstrate that the surfactants are incorporated into the growing crystals (which is a possible reason for crystal twinning) and can penetrate and adsorb within previously formed crystals. To characterize the interactions and the mass transfer of surfactants inside the crystalline protein matrices, we studied the infusion of lysozyme crystals with pyrene-based fluorescent surfactants by quantitative fluorescence microscopy. The fluorescence intensity data were fitted to the diffusion equation to obtain the effective diffusion coefficient of the fluorophore in the crystals. The diffusion coefficients obtained range from 2 × 10 -10 to 30 × 10 -10 cm 2 /s, depending on the type and size of the surfactant. The slow infusion is a consequence of the strong surfactant adsorption on the protein lattice; the estimated energy of adsorption is 10-12 kT. Fluorescence recovery after photobleaching (FRAP) experiments with saturated crystals showed that surfactant self-diffusion in these crystals is negligible, which is yet another indication of strong adsorption. The data shed light on the interactions between condensed protein phases and surfactants and are relevant to protein purification and crystallization and to the manipulation of proteins by light.

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Section: Introductionmentioning

confidence: 96%

Surfactant Diffusion into Lysozyme Crystal Matrices Investigated by Quantitative Fluorescence Microscopy

2000

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“…Uses of non-ionic surfactants in the cubic phase method shows the crystal growth to be more rapidly at lower pH than higher pH as seen with HEWL crystallization. Crystallization shows few membrane proteins but none of the membrane protein produced crystals which can be suitable for X-ray diffraction in contrast crystals were looks more suitable for soluble proteins (Mustafa et al, 1998). OJBS Science Publications…”

Section: Non-ionic Surfactants Approachmentioning

confidence: 99%

Different Approaches to Crystallization of Membrane Proteins

Doiphode¹,

Singh²

2012

OnLine Journal of Biological Sciences

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Crystallography is more like an art than science. Crystallizing membrane proteins are a big challenge; membrane proteins are present in the cell membrane and serve as cell support. The most important feature of membrane protein is that it contains both hydrophobic and hydrophilic regions on its surface. They are generally much more difficult to study than soluble proteins. The problem becomes more difficult when trying to obtain crystals to determine the high resolution structures of membrane proteins. We want to utilize this opportunity to briefly examine various approaches for crystallization of membrane proteins. The important factors for determining the success of crystallization experiments for membrane proteins lies in the purification, preparation of membrane samples, the environment in which the crystals are grown and the technique used to grow the crystals. All the X-ray structures of membrane protein are grown from preparations of detergents by different methods developed to crystallize. In this review different techniques for the crystallization of membrane proteins are being described. The cubic phase method also known as in meso method is discussed along with other methods to understand about the crystallization of membrane proteins, its general applicability, salt, detergent and screening effects on crystallization. Low volumes as nano-liter of samples can be used for crystallization. The effects of different detergents on the crystallization of membrane protein, as well as the use of surfactants like polyoxyethylene. Approach based on the detergent complexation to prove the ability of cyclodextrins to remove detergent from ternary mixtures in order to get 2D crystals. Crystallization of membrane proteins using non-ionic surfactants as well as Lipidic sponge phase and with swollen lipidic mesophases is discussed to better understand the crystallization of membrane proteins

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“…As soluble proteins share, to some extent, the aggregation problems of the membrane proteins, the mild nonionic surfactant may also be of use in crystallizing them. A few groups have successfully crystallized soluble proteins with nonionic surfactants (3,4). However, the mechanism of the soluble protein crystallization in the presence of nonionic surfactant is not well understood.…”

Section: Introductionmentioning

confidence: 99%

“…Variations with C 8 E 4 concentration of parameters obtained from SLS and DLS according to Eqs [1][2][3][4][5]. …”

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confidence: 99%

Investigation on the Mechanism of Crystallization of Soluble Protein in the Presence of Nonionic Surfactant

Jia

Narayanan

et al. 2005

Biophysical Journal

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The mechanism of crystallization of soluble, globular protein (lysozyme) in the presence of nonionic surfactant C8E4 (tetraoxyethylene glycol monooctyl ether) was examined using both static and dynamic light scattering. The interprotein interaction was found to be attractive in solution conditions that yielded crystals and repulsive in the noncrystallizing solution conditions. The validity of the second virial coefficient as a criterion for predicting protein crystallization could be established even in the presence of nonionic surfactants. Our experiments indicate that the origin of the change in interactions can be attributed to the adsorption of nonionic surfactant monomers on soluble proteins, which is generally assumed to be the case with only membrane proteins. This adsorption screens the hydrophobic attractive force and enhances the hydration and electrostatic repulsive forces between protein molecules. Thus at low surfactant concentration, the effective protein-protein interaction remains repulsive. Large surfactant concentrations promote protein crystallization, possibly due to the attractive depletion force caused by the intervening free surfactant micelles.

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A novel approach for the crystallization of soluble proteins using non-ionic surfactants

Cited by 6 publications

References 16 publications

Surfactant Diffusion into Lysozyme Crystal Matrices Investigated by Quantitative Fluorescence Microscopy

Surfactant Diffusion into Lysozyme Crystal Matrices Investigated by Quantitative Fluorescence Microscopy

Different Approaches to Crystallization of Membrane Proteins

Investigation on the Mechanism of Crystallization of Soluble Protein in the Presence of Nonionic Surfactant

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