Combining thermostable mutations increases the stability of λ repressor

Robert S. Stearman, Alan D. Frankel, Ernesto I Freire, Beishan Liu, Carl O. Pabo

Research output: Contribution to journalArticle

Abstract

We have combined three mutations previously shown to stabilize λ repressor against thermal denaturation. Two of these mutations are in helix 3, where Gly-46 and Gly-48 have been replaced by alanines [Hecht, M. H., et al. (1986) Proteins: Struct., Funct., Genet. I, 43-46]. The other mutation, which replaces Tyr-88 with cysteine, allows the protein to form an intersubunit disulfide bond [Sauer, R. T., et al. (1986) Biochemistry 25, 5992-5998]. Calorimetric measurements show that the two alanine substitutions stabilize repressor by about 8°C, that the disulfide bond stabilizes repressor by about 8°C, and that the triple mutant is 16°C more stable than wild-type repressor.

Original languageEnglish (US)
Pages (from-to)7571-7574
Number of pages4
JournalBiochemistry®
Volume27
Issue number19
StatePublished - 1988

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Disulfides
Alanine
Mutation
Biochemistry
Denaturation
Cysteine
Viverridae
Proteins
Substitution reactions
Hot Temperature

ASJC Scopus subject areas

  • Biochemistry

Cite this

Stearman, R. S., Frankel, A. D., Freire, E. I., Liu, B., & Pabo, C. O. (1988). Combining thermostable mutations increases the stability of λ repressor. Biochemistry®, 27(19), 7571-7574.

Combining thermostable mutations increases the stability of λ repressor. / Stearman, Robert S.; Frankel, Alan D.; Freire, Ernesto I; Liu, Beishan; Pabo, Carl O.

In: Biochemistry®, Vol. 27, No. 19, 1988, p. 7571-7574.

Research output: Contribution to journalArticle

Stearman, RS, Frankel, AD, Freire, EI, Liu, B & Pabo, CO 1988, 'Combining thermostable mutations increases the stability of λ repressor', Biochemistry®, vol. 27, no. 19, pp. 7571-7574.
Stearman RS, Frankel AD, Freire EI, Liu B, Pabo CO. Combining thermostable mutations increases the stability of λ repressor. Biochemistry®. 1988;27(19):7571-7574.
Stearman, Robert S. ; Frankel, Alan D. ; Freire, Ernesto I ; Liu, Beishan ; Pabo, Carl O. / Combining thermostable mutations increases the stability of λ repressor. In: Biochemistry®. 1988 ; Vol. 27, No. 19. pp. 7571-7574.
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