A spectroscopic and equilibrium binding analysis of cationic detergent-protein interactions using soluble and insoluble recombinant porcine growth hormone

Cardamone, Michael; Puri, N K; Sawyer, William H.; Capon, Robert J.; Brandon, Malcolm R.

doi:10.1016/0167-4838(94)90074-4

Cited by 14 publications

(8 citation statements)

References 24 publications

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“…The trend in G6PD stability observed in Figure 3 can be understood based on the mechanism of protein denaturation induced by ionic surfactants. It is generally accepted that the denaturation of proteins by ionic surfactants results from a combination of electrostatic and hydrophobic interactions (Cardamone et al, 1994). In an aqueous solution containing protein and ionic surfactant, the ionic surfactant monomers first bind electrostatically to oppositely charged residues at the protein surface (site-specific binding), and induce an expansion of the protein structure.…”

Section: G6pd Partitioning In the C 10 E 4 /C N Tab/buffer Systemsmentioning

confidence: 99%

Glucose‐6‐phosphate dehydrogenase partitioning in two‐phase aqueous mixed (nonionic/cationic) micellar systems

Rangel-Yagui

Lam

Kamei

et al. 2003

Biotech & Bioengineering

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The enzyme glucose-6-phosphate dehydrogenase (G6PD) plays an important role in maintaining the level of NADPH and in producing pentose phosphates for nucleotide biosynthesis. It is also of great value as an analytical reagent, being used in various quantitative assays. In searching for new strategies to purify this enzyme, the partitioning of G6PD in two-phase aqueous mixed (nonionic/cationic) micellar systems was investigated both experimentally and theoretically. Our results indicate that the use of a two-phase aqueous mixed micellar system composed of the nonionic surfactant C(10)E(4) (n-decyl tetra(ethylene oxide)) and the cationic surfactant C(n)TAB (alkyltrimethylammonium bromide, n = 8, 10, or 12) can improve significantly the partitioning behavior of G6PD relative to that obtained in the two-phase aqueous C(10)E(4) micellar system. This improvement can be attributed to electrostatic attractions between the positively charged mixed (nonionic/cationic) micelles and the net negatively charged enzyme G6PD, resulting in the preferential partitioning of G6PD to the top, mixed micelle-rich phase of the two-phase aqueous mixed micellar systems. The effect of varying the cationic surfactant tail length (n = 8, 10, and 12) on the denaturation and partitioning behavior of G6PD in the C(10)E(4) /C(n)TAB/buffer system was investigated. It was found that C(8)TAB is the least denaturing to G6PD, followed by C(10)TAB and C(12)TAB. However, the C(10)E(4)/C(12)TAB/buffer system generated stronger electrostatic attractions with the net negatively charged enzyme G6PD than the C(10)E(4)/C(10)TAB/buffer and the C(10)E(4)/C(8)TAB/buffer systems, when using the same amount of cationic surfactant. Overall, the two-phase aqueous mixed (C(10)E(4)/C(10)TAB) micellar system yielded the highest G6PD partition coefficient of 7.7, with a G6PD yield in the top phase of 71%, providing the optimal balance between the denaturing effect and the electrostatic attractions for the three cationic surfactants examined. A recently developed theoretical framework to predict protein partition coefficients in two-phase aqueous mixed (nonionic/ionic) micellar systems was implemented, and the theoretically predicted G6PD partition coefficients were found to be in reasonable quantitative agreement with the experimentally measured ones.

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Section: G6pd Partitioning In the C 10 E 4 /C N Tab/buffer Systemsmentioning

confidence: 99%

Glucose‐6‐phosphate dehydrogenase partitioning in two‐phase aqueous mixed (nonionic/cationic) micellar systems

Rangel-Yagui

Lam

Kamei

et al. 2003

Biotech & Bioengineering

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“…Although it is well known that ionic surfactants can bind to proteins and induce denaturation, this effect has been shown to depend on the type and concentration of the ionic surfactant (Cardamone et al, 1994;Gelamo and Tabak, 2000). Therefore, when working with a two-phase aqueous mixed (nonionic/ionic) micellar system, the amount of ionic surfactant added should be sufficiently high to induce a significant change in the protein partitioning behavior, but sufficiently low to prevent severe denaturation of the protein (Kamei et al, 2002a,b).…”

Section: Spherical Micelle Cylindrical Micelle Bilayermentioning

confidence: 99%

Two-phase aqueous micellar systems: an alternative method for protein purification

Rangel-Yagui

Pessoa

Blankschtein

2004

Braz. J. Chem. Eng.

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-Two-phase aqueous micellar systems can be exploited in separation science for the extraction/purification of desired biomolecules. This article reviews recent experimental and theoretical work by Blankschtein and co-workers on the use of two-phase aqueous micellar systems for the separation of hydrophilic proteins. The experimental partitioning behavior of the enzyme glucose-6-phosphate dehydrogenase (G6PD) in two-phase aqueous micellar systems is also reviewed and new results are presented. Specifically, we discuss very recent work on the purification of G6PD using: i) a two-phase aqueous micellar system composed of the nonionic surfactant n-decyl tetra(ethylene oxide) (C 10 E 4 ), and (ii) a two-phase aqueous mixed micellar system composed of C 10 E 4 and the cationic surfactant decyltrimethylammonium bromide (C 10 TAB). Our results indicate that the two-phase aqueous mixed (C 10 E 4 /C 10 TAB) micellar system can improve significantly the partitioning behavior of G6PD relative to that observed in the two-phase aqueous C 10 E 4 micellar system.

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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.…”

mentioning

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

102

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A spectroscopic and equilibrium binding analysis of cationic detergent-protein interactions using soluble and insoluble recombinant porcine growth hormone

Cited by 14 publications

References 24 publications

Glucose‐6‐phosphate dehydrogenase partitioning in two‐phase aqueous mixed (nonionic/cationic) micellar systems

Glucose‐6‐phosphate dehydrogenase partitioning in two‐phase aqueous mixed (nonionic/cationic) micellar systems

Two-phase aqueous micellar systems: an alternative method for protein purification

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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