Combining thermostable mutations increases the stability of lambda repressor.

Abstract:

:We have combined three mutations previously shown to stabilize lambda 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 Two of these mutations are in helix 3, where Gly-46 and Gly-48 have been replaced by alanines [Hecht, M. H., et al.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. 1, 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 degrees C, that the disulfide bond stabilizes repressor by about 8 degrees C, and that the triple mutant is 16 degrees C more stable than wild-type repressor.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Stearman RS,Frankel AD,Freire E,Liu BS,Pabo CO

doi

10.1021/bi00419a059

subject

Has Abstract

pub_date

1988-09-20 00:00:00

pages

7571-4

issue

19

eissn

0006-2960

issn

1520-4995

journal_volume

27

pub_type

杂志文章