Protein refolding in predominantly organic media markedly enhanced by common salts

Rariy, Roman V.; Klibanov, Alexander M.

doi:10.1002/(sici)1097-0290(19990320)62:6<704::aid-bit9>3.0.co;2-s

Cited by 33 publications

(7 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is worth noting that no correlation has been observed between the conformational state of lysozyme dissolved in the six solvents and their physicochemical characteristics as dielectric constant, dipole moment, empirical solvent polarity parameter (E T N ), and Hildebrand solubility parameter. The results of the present study are consistent with the lack of lysozyme refolding/reoxidation in 99% ethylene glycol, methanol, DMSO, and formamide (Rariy and Klibanov, 1999), which is in contrast to the correct refolding/ reoxidation observed in 99% glycerol (Rariy and Klibanov, 1997).…”

Section: Comparison Of the Nmr And CD Resultssupporting

confidence: 89%

Structure of lysozyme dissolved in neat organic solvents as assessed by NMR and CD spectroscopies

Knubovets

Osterhout²,

Klibanov

1999

Biotechnol. Bioeng.

Self Cite

View full text Add to dashboard Cite

The structure of the model protein hen egg‐white lysozyme dissolved in water and in five neat organic solvents (ethylene glycol, methanol, dimethylsulfoxide (DMSO), formamide, and dimethylformamide (DMF)) has been examined by means of 1H NMR and circular dichroism (CD) spectroscopies. The NMR spectra of lysozyme reveal the lack of a defined tertiary structure in all five organic solvents, although the examination of line widths suggests the possibility of some ordered structure in ethylene glycol and in methanol. The near‐UV CD spectra of the protein suggest no tertiary structure in lysozyme dissolved in DMSO, formamide, and DMF, while a distinctive (albeit less pronounced than in water) tertiary structure is seen in ethylene glycol and a drastically changed one in methanol. A highly developed secondary structure was observed by far‐UV CD in ethylene glycol and methanol; interestingly, the α‐helix content of the protein in both was greater than in water, while the β‐structure content was lower. (Solvent absorbance in the far‐UV region prevents conclusions about the secondary structure in DMSO, formamide and DMF.) © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 63: 242–248, 19999.

show abstract

Section: Comparison Of the Nmr And CD Resultssupporting

confidence: 89%

Structure of lysozyme dissolved in neat organic solvents as assessed by NMR and CD spectroscopies

Knubovets

Osterhout²,

Klibanov

1999

Biotechnol. Bioeng.

Self Cite

View full text Add to dashboard Cite

show abstract

“…As discussed above, polyamines effectively suppressed heat-induced aggregation; thus the amino group may play a key role in preventing heat-induced aggregation. It is worth noting that salts increase the solubility of aggregation-prone protein in the presence of organic solvent and amphiphilic polymer [117,118].…”

Section: Saltsmentioning

confidence: 99%

Effect of Additives on Protein Aggregation

Hamada

Arakawa²,

Shiraki

2009

CPB

222

131

View full text Add to dashboard Cite

This paper overviews solution additives that affect protein stability and aggregation during refolding, heating, and freezing processes. Solution additives are mainly grouped into two classes, i.e., protein denaturants and stabilizers. The former includes guanidine, urea, strong ionic detergents, and certain chaotropic salts; the latter includes certain amino acids, sugars, polyhydric alcohols, osmolytes, and kosmotropic salts. However, there are solution additives that are not unambiguously placed into these two classes, including arginine, certain divalent cation salts (e.g., MgCl(2)) and certain polyhydric alcohols (e.g., ethylene glycol). Certain non-ionic or non-detergent surfactants, ionic liquids, amino acid derivatives, polyamines, and certain amphiphilic polymers may belong to this class. They have marginal effects on protein structure and stability, but are able to disrupt protein interactions. Information on additives that do not catalyze chemical reactions nor affect protein functions helps us to design protein solutions for increased stability or reduced aggregation.

show abstract

“…The interest for the effects of these cosolvents on conformational and dynamic properties of proteins is more general. For example, these cosolvents are used in studying protein folding and also used as additive to stabilize protein structure during crystallization (Sawano et al, 1992;Rariy and Klibanov, 1998;Sousa, 1995;Farnum and Zukoski, 1999;Rubinson et al, 2000). However their role in these processes is poorly understood.…”

Section: Introductionmentioning

confidence: 99%

Dielectric Behavior of Lysozyme and Ferricytochrome-c in Water/Ethylene-Glycol Solutions

Bonincontro

Cinelli

Onori

et al. 2004

Biophysical Journal

View full text Add to dashboard Cite

This work deals with a dielectric study at radio frequencies of the influence at room temperature of two organic molecules, known as cryo-protectants, ethylene-glycol and glycerol, on conformational and dynamic properties of two model proteins, lysozyme (lys) from chicken egg-white and ferricytochrome-c (cyt-c) from horse heart. Cyt-c is a compact globular protein whereas lys is composed of two structural domains, separated by the active site cleft. Measurements were carried out at the fixed temperature of 20 degrees C varying the concentration of the cosolvent up to 90% w/w. From the analysis of the dielectric relaxation of the protein solution, the effective hydrodynamic radius and the electric dipole moment of the protein were calculated as a function of the cosolvent concentration. The data show that glycerol does not modify significantly the conformation of both proteins and cyt-c is also stable in the presence of ethylene-glycol. On the contrary ethylene-glycol strongly affects the dielectric response of lysozyme denoting a specific effect on its conformation and dynamics. The data are coherently interpreted hypothesizing that glycol molecule wedges between and separates the two domains of lys making them rotationally independent.

show abstract

Protein refolding in predominantly organic media markedly enhanced by common salts

Cited by 33 publications

References 24 publications

Structure of lysozyme dissolved in neat organic solvents as assessed by NMR and CD spectroscopies

Structure of lysozyme dissolved in neat organic solvents as assessed by NMR and CD spectroscopies

Effect of Additives on Protein Aggregation

Dielectric Behavior of Lysozyme and Ferricytochrome-c in Water/Ethylene-Glycol Solutions

Contact Info

Product

Resources

About