Protein Engineering vol. 9 no. 3 pp. 315-316, 1996
© 1996 Oxford University Press
LETTER |
On the choice of reference mutant states in the application of the double-mutant cycle method
Department of Structural Biology, Weizmann Institute Rehovot 76100, Israel
1To whom correspondence should be addressed
Pairwise interactions in proteins can be detected and, in certain circumstances, their strength measured by applying the method of double-mutant cycles. Such cycles comprise wild type protein, two single mutants and the corresponding double mutant The analysis of double-mutant cycles is most straightforward when the mutations are to alanine since interactions are mostly removed without new interactions being formed. Here, ‘not-to-alanine’ double-mutant cycles are analysed. It is shown that a ‘not-to-alanine’ double-mutant cycle can be decomposed into four double-mutant cycles with mutations only to alanine. The coupling energy corresponding to the ‘not-to-alanine’ double-mutant cycle is expressed as a function of the coupling energies of these four cycles.
Keywords: double-mutant cycles/mutagenesis/protein engineering
Received October 24, 1995; accepted December 21, 1995.
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