Determination of the Binding Isotherm and Size of the Bovine Serum Albumin-Sodium Dodecyl Sulfate Complex by Diffusion-Ordered 2D NMR

Chen, Aidi; Wu, Donghui; Johnson, Charles S. Jr.

doi:10.1021/j100002a054

Cited by 108 publications

(100 citation statements)

References 8 publications

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“…In addition, the diffusion coefficient of acetic acid also present in the sample was measured and used as a reference 19 in order to compare diffusion coefficients of samples at different concentrations and buffer conditions. The experimentally measured translational diffusion coefficients were corrected using the following relationship:…”

Section: Translational Diffusion Measurementsmentioning

confidence: 99%

Merozoite surface protein 2 of Plasmodium falciparum: Expression, structure, dynamics, and fibril formation of the conserved N‐terminal domain

et al. 2007

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Merozoite surface protein 2 (MSP2) is a GPI-anchored protein on the surface of the merozoite stage of the malaria parasite Plasmodium falciparum. It is largely disordered in solution, but has a propensity to form amyloid-like fibrils under physiological conditions. The N-terminal conserved region (MSP2(1-25)) is part of the protease-resistant core of these fibrils. To investigate the structure and dynamics of this region, its ability to form fibrils, and the role of individual residues in these properties, we have developed a bacterial expression system that yields > or =10 mg of unlabeled or (15)N-labeled peptide per litre of culture. Two recombinant versions of MSP2(1-25), wild-type and a Y7A/Y16A mutant, have been produced. Detailed conformational analysis of the wild-type peptide and backbone (15)N relaxation data indicated that it contains beta-turn and nascent helical structures in the central and C-terminal regions. Residues 6-21 represent the most ordered region of the structure, although there is some flexibility around residues 8 and 9. The 10-residue sequence (MSP2(7-16)) (with two Tyr residues) was predicted to have a higher propensity for beta-aggregation than the 8-mer sequence (MSP2(8-15)), but there was no significant difference in conformation between MSP2(1-25) and [Y7A,Y16A]MSP2(1-25) and the rate of fibril formation was only slightly slower in the mutant. The peptide expression system described here will facilitate further mutational analyses to define the roles of individual residues in transient structural elements and fibril formation, and thus contribute to the further development of MSP2 as a malaria vaccine candidate.

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Section: Translational Diffusion Measurementsmentioning

confidence: 99%

Merozoite surface protein 2 of Plasmodium falciparum: Expression, structure, dynamics, and fibril formation of the conserved N‐terminal domain

et al. 2007

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“…The denaturation process apparently destroys the primary binding sites for SDS and detergent attachment becomes purely hydrophobic [12]. The transformation of the protein towards the random coil structure increases the Stokes'-Einstein radius [19,54]. This region extends until the SDS saturation point (polymer saturation point = psp) is reached [21,27].…”

Section: Binding Isothermsmentioning

confidence: 99%

Detection of coexisting protein conformations in capillary zone electrophoresis subsequent to transient contact with sodium dodecyl sulfate solutions

et al. 2005

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Non-native conformations of proteins were generated by temporary contact with aqueous solutions of sodium dodecyl sulfate (SDS) and separated from the native state with capillary zone electrophoresis (CZE) in alkaline borate buffer deficient of SDS. Nine proteins at concentrations of 2.0 or 3.0 mg.L(-1) were compared in terms of their susceptibility to SDS. For superoxide dismutase and ferritin the tendency of unfolding was modest with < 25% of the protein being transformed to the non-native state at 10 mmol.L(-1) SDS. Highest susceptibility was observed for albumin, myoglobin (Mb), and hemoglobin with > 75% in the non-native state even at 2.0 mmol.L(-1) SDS. The influence of varying SDS concentrations on the conformational state of Mb was tested. Increasing the SDS concentration, circular dichroism revealed a reduction in alpha-helix, an increase in random coil, and an introduction of beta-sheet, which is absent in native structure. Modifications in the secondary structure were in agreement with distinct changes in the shape of the non-native Mb peak in CZE and make a gradual unfolding/refolding process with several coexisting molten globules instead of two-state transition of conformations most plausible for Mb. CZE was found to contribute to a further understanding of holo-Mb transformation towards a population of non-native conformations (i) by means of calculated peak area ratios of native to non-native states, which showed sigmoid transition, (ii) by detecting the release of the prosthetic heme group, and (iii) by changes in the effective electrophoretic mobility of the Mb-SDS peaks. Reconstituted holo-Mb forms differed in the Soret band around 410 nm, indicating diversity in the conformation of the heme pocket.

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“…Especially for the prevention of physical degradation, surface active agents like detergents are often necessary [17,18]. Therefore, a number of biophysical techniques are used to investigate and understand protein-excipient interactions [6,[23][24][25][26][27][28][29][30]. Detergents are amphiphilic in nature, containing a hydrophilic head group and a hydrophobic part, and this dual property causes detergents to adopt a specific orientation at interfaces and in aqueous solutions.…”

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confidence: 99%

A thermodynamic analysis of the binding interaction between polysorbate 20 and 80 with human serum albumins and immunoglobulins: A contribution to understand colloidal protein stabilisation

Garidel

Hoffmann

Blume

2009

Biophysical Chemistry

106

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Determination of the Binding Isotherm and Size of the Bovine Serum Albumin-Sodium Dodecyl Sulfate Complex by Diffusion-Ordered 2D NMR

Cited by 108 publications

References 8 publications

Merozoite surface protein 2 of Plasmodium falciparum: Expression, structure, dynamics, and fibril formation of the conserved N‐terminal domain

Merozoite surface protein 2 of Plasmodium falciparum: Expression, structure, dynamics, and fibril formation of the conserved N‐terminal domain

Detection of coexisting protein conformations in capillary zone electrophoresis subsequent to transient contact with sodium dodecyl sulfate solutions

A thermodynamic analysis of the binding interaction between polysorbate 20 and 80 with human serum albumins and immunoglobulins: A contribution to understand colloidal protein stabilisation

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