Non-detergent sulphobetaines: a new class of molecules that facilitate in vitro protein renaturation.
BACKGROUND
Attempts to renature proteins often yield aggregates rather than native protein. To minimize aggregation, low protein concentrations and/or solubilizing agents are used. Here, we test new solubilizing molecules, non-detergent sulphobetaines, to improve the renaturation of two very different enzymes, hen egg white lysozyme and bacterial beta-D-galactosidase.


RESULTS
The renaturation was conducted in the presence of five different sulphobetaines and the yield of active enzyme was measured. The five sulphobetaines improved the yield of native lysozyme up to 12-fold. Some sulphobetaines improved the yield of galactosidase up to 80-fold, but one reduced it 100-fold.


CONCLUSIONS
Non-detergent sulphobetaines strongly affect the balance between aggregation and folding. Their effect depends on their structure and on their interactions with folding intermediates. These results should serve as a basis for designing more efficient sulphobetaines; for designing improved renaturation protocols using existing sulphobetaines; and for characterizing folding intermediates that interact with sulphobetaines.
Goldberg
M E
ME
Unité de Biochimie Cellulaire (CNRS URA 1129), Institute Pasteur, Paris, France. goldberg@pasteur.fr
Expert-Bezançon
N
N
Vuillard
L
L
Rabilloud
T
T
D000818:Animals; D001622:Betaine / Q000031:analogs & derivatives* / Q000494:pharmacology; D002645:Chickens; D004926:Escherichia coli / Q000201:enzymology; D005260:Female; D066298:In Vitro Techniques; D009113:Muramidase / Q000737:chemistry; D017510:Protein Folding; D001616:beta-Galactosidase / Q000737:chemistry
D001622:Betaine; D009113:Muramidase; D001616:beta-Galactosidase