Functional consequences of single:double ring transitions in chaperonins: life in the cold

Molecular Microbiology
Manuel FerrerPeter N Golyshin

Abstract

The cpn60 and cpn10 genes from psychrophilic bacterium, Oleispira antarctica RB8, showed a positive effect in Escherichia coli growth at low temperature, shifting its theoretical minimal growth temperature from +7.5 degrees C to -13.7 degrees C [Ferrer, M., Chernikova, T.N., Yakimov, M., Golyshin, P.N., and Timmis, K.N. (2003) Nature Biotechnol 21: 1266-1267]. To provide experimental support for this finding, Cpn60 and 10 were overproduced in E. coli and purified to apparent homogeneity. Recombinant O.Cpn60 was identical to the native protein based on tetradecameric structure, and it dissociates during native PAGE. Gel filtration and native PAGE revealed that, in vivo and in vitro, (O.Cpn60)(7) was the active oligomer at 4-10 degrees C, whereas at > 10 degrees C, this complex was converted to (O.Cpn60)(14). The dissociation reduces the ATP consumption (energy-saving mechanism) and increases the refolding capacity at low temperatures. In order for this transition to occur, we demonstrated that K468 and S471 may play a key role in conforming the more advantageous oligomeric state in O.Cpn60. We have proved this hypothesis by showing that single and double mutations in K468 and S471 for T and G, as in E.GroEL, produced a more stab...Continue Reading

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Citations

Sep 22, 2012·Extremophiles : Life Under Extreme Conditions·Seiji YamauchiHidenori Hayashi
Oct 7, 2009·Proceedings of the National Academy of Sciences of the United States of America·Bong-Gyoon HanRobert M Glaeser
Aug 6, 2004·Applied and Environmental Microbiology·Manuel FerrerPeter N Golyshin
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Aug 11, 2021·MSystems·Tomás Alarcón-SchumacherBeatriz Díez

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