Comparison of denaturation by guanidine hydrochloride of the wild type tryptophan synthase alpha-subunit of Escherichia coli and two mutant protein (Glu 49 replaced by Met or Gln).


Abstract

In order to elucidate the roles of individual amino acid residues in the conformational stability of proteins, the denaturation by guanidine hydrochloride of the wild-type trytophan synthase alpha-subunit of Escherichia coli and two mutant proteins, trpA33 (Glu 49 leads to Met) and trpA11 (Glu 49 leads to Gln), has been compared by means of CD measurements at pH 7.0 and various temperatures. CD spectra of the two mutant proteins were similar to that of the wild-type protein. The trpA33 and the trpA11 proteins were more and less resistant, respectively, to guanidine hydrochloride than the wild-type protein at 9.7 to 49.6 degrees C. The free energy change of unfolding in water delta delta Gnd H2O, was evaluated assuming a three state denaturation, since the denaturation curves of three proteins suggested the presence of one stable intermediate. The values of delta Gnd H2O for the trpA33, the wild-type, and the trpA11 proteins at 25.8 degrees C and pH 7.0 were 13.4,8.8, and 6.3 kcal/mol, respectively. The delta Gnd H2O of the trpA11 protein was almost independent of temperature, though that of the trpA33 protein was remarkably dependent on temperature. The conformation stabilities of the three proteins were correlated with the hydrophobicities of the substituted amino acid residues. Study holds ProTherm entries: 2476, 2477, 2478 Extra Details: additive : EDTA(0.1 mM), conformational stability; hydrophobicity; Escherichia coli;,alpha subunits of tryptophan synthase; free energy

Submission Details

ID: 6estGvSF4

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:19 p.m.

Version: 1

Publication Details
Yutani K;Ogasahara K;Suzuki M;Sugino Y,J. Biochem. (1979) Comparison of denaturation by guanidine hydrochloride of the wild type tryptophan synthase alpha-subunit of Escherichia coli and two mutant protein (Glu 49 replaced by Met or Gln). PMID:378988
Additional Information

Sequence Assay Result Units