Conformational Transitions Provoked by Organic Solvents in β-lactoglobulin: Can a Molten Globule Like Intermediate Be Induced by the Decrease in Dielectric Constant?
Intermediate States, Molten Globule, Organic Solvents, Protein Folding, 8-ANS, Time-resolved Fluorescence
Digital Object Identifier (DOI)
Background: It is known that nonnative states of protein molecules can exist in living cells and can be involved in a number of physiological processes. It has also been established that the membrane surface can be responsible for the partial denaturation of proteins due to negative charges on it. The local decrease in the effective dielectric constant of water near the organic surface has been suggested to be an additional driving force for protein denaturation in the membrane field, but data to confirm this suggestion were lacking.
Results: Conformational transitions induced in β-lactoglobulin by methanol, ethanol, isopropanol, dimethylformamide and dioxane were studied by near and far UV circular dichroism, steady-state tryptophan fluorescence and fluorescence decay of 8-anilinonaphthalene-1-sulfonate (8-ANS). The existence of at least two noncoinciding cooperative transitions has been established in all solvent systems studied. The first of these transitions describes the disruption of rigid tertiary structure in protein molecules, while the second reflects the formation of an expanded helical conformation typical of proteins in concentrated organic solvents. This means that the organic solvents provoke the formation of a denatured intermediate state with pronounced secondary structure and native-like compactness. We show that the positions of maxima in fI versus dielectric constant dependence virtually coincide for all five solvent systems studied.
Conclusions: The decrease in the dielectric constant of the solvent induces in β-lactoglobulin an equilibrium intermediate state. This state, being denatured, is relatively compact and has pronounced secondary structure and high affinity for the hydrophobic fluorescent probe 8-ANS, i.e. possesses all the properties of the molten globule intermediate state.
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Citation / Publisher Attribution
Folding and Design, v. 2, issue 3, p. 163-172
Scholar Commons Citation
Uversky, Vladimir N.; Narizhneva, Natalya V.; Kirschstein, Steffen Omar; Winter, Stefan; and Löber, Günter, "Conformational Transitions Provoked by Organic Solvents in β-lactoglobulin: Can a Molten Globule Like Intermediate Be Induced by the Decrease in Dielectric Constant?" (1997). Molecular Medicine Faculty Publications. 641.