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J. Biol. Chem., Vol. 262, Issue 10, 4486-4491, Apr, 1987
S Tandon and PM Horowitz
We have established the generality of using detergents for facilitating the
reactivation of 6 M guanidinium chloride-denatured rhodanese that was
recently described for the nonionic detergent lauryl maltoside (LM)
(Tandon, S., and Horowitz, P. (1986) J. Biol. Chem. 261, 15615-15618). We
report here that not only LM but other nonionic as well as ionic and
zwitterionic detergents also have favorable effects in reactivating the
denatured enzyme. Not all detergents are useful, and the favorable effects
occur over a limited concentration range. Above and below that range there
is little or no effect. Zwittergents, which represent a homologous series
with varying critical micelle concentrations (CMCs) are effective only
above their CMCs. Induction phases occur in the progress curves of
rhodanese refolded in the presence of the effective detergents, suggesting
the presence of refolding intermediates that are apparently stabilized by
detergent interactions. Gel filtration chromatography of rhodanese with and
without LM suggests that even though the renaturation of the denatured
enzyme requires detergent at concentrations above its CMC, the enzyme does
not bind an amount of detergent equivalent to a micelle. It is suggested
that renaturation of other proteins might also be assisted by inclusion of
"nondenaturing" detergents, although the optimal conditions will have to be
determined for each individual case.
Detergent-assisted refolding of guanidinium chloride-denatured rhodanese. The effects of the concentration and type of detergent
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