Fractionation techniques in a hydro-organic environment. I. Sulfolane as a solvent for hydrophobic proteins |
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Authors: | G Vecchio P G Righetti M Zanoni G Artoni E Gianazza |
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Affiliation: | 1. Laboratorio di Chimica degli Ormoni del CNR, via Mario Bianco 9, Milano I-20133, Italy;2. Chair of Biochemistry, School of Pharmaceutical Sciences, University of Milano, via Celoria 2, Milano I-20133, Italy;3. Department of Biomedical Sciences and Technologies, University of Milano, via Celoria 2, Milano I-20133, Italy |
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Abstract: | Sulfolane (thiophene, tethrahydro-1,1-dioxide), at concentrations of 4 M or above, is an efficient solubilizing agent for water-insoluble proteins (e.g., zein or globin chains). In comparison with urea, it appears indefinitely stable in aqueous solutions and does not chemically modify proteins upon storage. Moreover, it favors protein structure, i.e., it increases their alpha-helix content, while urea decreases it. Sulfolane is compatible with electrophoretic techniques (it only slightly reduces polyacrylamide polymerization efficiency and it does not interfere with protein and peptide detection methods) and with chromatographic methods (it has negligible A280 nm). With hydrophilic proteins, sulfolane behaves as a mild denaturant and precipitates them at concentrations between 5 and 7 M. |
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Keywords: | sulfolane hydrophobic proteins urea protein denaturation electrophoresis ion-exchange chromatography |
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