Cooperativity mutants of bacteriophage lambda cI repressor: temperature dependence of self-assembly

Biochemistry
D S Burz, G K Ackers

Abstract

Analytical ultracentrifugation was used to study higher order self-assembly of lambda cI repressors, including eight mutants whose monomer to dimer reactions were recently characterized [Burz et al. (1994) Biochemistry 33, 8399]. Six of the mutants were found to remain dimeric up to 50 microM total protein; the remaining mutants (EK102 and PT158) were found to undergo higher order oligomerization, as does wild-type cI. For these three repressors, we determined the stoichiometries and energetics of higher order assembly over the temperature range 5-40 degrees C. Weak dimerization exhibited by two other mutants, SN228 and SR228, was also evaluated by sedimentation equilibrium over this same temperature range. The end-state for higher order assembly of wild-type cI was determined to be octameric, in agreement with Senear et al. [(1993) Biochemistry 32, 6179-6189]. The assembly free energies resolved by the sedimentation analysis program NONLIN [Johnson, M. L., et al. (1981) Biophys. J. 36, 575-588] leads to the prediction that tetramers may contribute significantly to the intermediate populations during assembly. This analysis of the species populations is in accord with recent conclusions from fluorescence anisotropy studies [Ban...Continue Reading

References

Mar 5, 1989·Journal of Molecular Biology·F S Gimble, R T Sauer
Jul 4, 1970·Nature·V PirrottaM Ptashne

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Citations

Feb 12, 2004·Genes & Development·Ian B DoddJ Barry Egan
Mar 31, 2005·Current Opinion in Genetics & Development·Jose M G Vilar, Leonor Saiz
Feb 1, 2008·Journal of Molecular Biology·Kim C GieseJohn W Little
Feb 18, 2005·Comptes rendus biologies·Michèle Amouyal
Oct 19, 2004·Journal of Theoretical Biology·Audun Bakk, Ralf Metzler
Dec 26, 2003·Biophysical Journal·Audun BakkKim Sneppen
Oct 17, 1998·Methods : a Companion to Methods in Enzymology·D F SenearT M Laue
Jun 2, 2021·Journal of the Royal Society, Interface·Pankaj Gautam, Sudipta Kumar Sinha

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