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. 1991 Sep 1;88(17):7715–7718. doi: 10.1073/pnas.88.17.7715

Cold denaturation and 2H2O stabilization of a staphylococcal nuclease mutant.

L C Antonino 1, R A Kautz 1, T Nakano 1, R O Fox 1, A L Fink 1
PMCID: PMC52373  PMID: 1652762

Abstract

Cold denaturation is now recognized as a general property of proteins but has been observed only under destabilizing conditions, such as moderate denaturant concentration or low pH. By destabilizing the protein using site-directed mutagenesis, we have observed cold denaturation at pH 7.0 in the absence of denaturants in a mutant of staphylococcal nuclease, which we call NCA S28G for a hybrid protein between staphylococcal nuclease and concanavalin A in which there is the point mutation Ser-28----Gly. The temperature of maximum stability (tmax) as determined by circular dichroism (CD) was 18.1 degrees C, and the midpoints of the thermal unfolding transitions (tm) were 0.6 degrees C and 30.0 degrees C. These values may be compared with the tm of 52.5 degrees C for wild-type staphylococcal nuclease, for which no cold denaturation was observed under these conditions. When the stability of the mutant was examined in 2H2O by NMR, CD, or fluorescence, a substantial increase in the amount of folded protein at the tmax was noted as well as a decrease in tmax, reflecting increased stability.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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