Document Detail

Effect of single amino acid substitutions at positions 49 and 60 on the thermal unfolding of the tryptophan synthase alpha subunit from Salmonella typhimurium.
MedLine Citation:
PMID:  1989494     Owner:  NLM     Status:  MEDLINE    
We have used circular dichroism measurements to compare the thermal unfolding of the wild type tryptophan synthase alpha subunit from Salmonella typhimurium with that of seven mutant forms with single amino acid replacements at two active site residues. Glutamic acid 49 has been replaced by phenylalanine, glutamine, or aspartic acid. Aspartic acid 60 has been replaced by alanine, aspartic acid, asparagine, or tyrosine. Thermodynamic properties (delta G, delta H, delta S, and Tm) of the wild type and mutant forms have been determined experimentally by measuring the free energy of unfolding as a function of temperature. Increasing the pH from 7.0 to 8.8 decreases the tm of the wild type alpha subunit from 56 to 45 degrees C. The thermal unfolding of the wild type alpha subunit and of six of the seven mutant forms can be described as reversible, two-state transitions. In contrast, the melting curve of a mutant alpha subunit in which aspartic acid 60 is replaced by tyrosine indicates the presence of a folding intermediate which may correspond to a "molten globule." Correlations between our observations and previous folding studies and the X-ray crystallographic structure are presented. Substitution of glutamic acid 49, which is located in the hydrophobic "pit" of an eight-fold alpha/beta barrel, by a hydrophobic phenylalanine residue increases the tm from 56 to 60 degrees C. In contrast, replacement of aspartic acid 60, which is accessible to solvent, results in small reductions in the thermal stability.
H Kanzaki; P McPhie; E W Miles
Related Documents :
8572694 - Structure and function of fas-1a, a gene encoding a putative fatty acid synthetase dire...
17158734 - Mutations in alternative carbon utilization pathways in candida albicans attenuate viru...
15866214 - Mutational analysis of the helicobacter pylori carbonic anhydrases.
697764 - The lipid composition of a barley mutant lacking chlorophyll b.
21644574 - Polycarbonates from the polyhydroxy natural product quinic acid.
486394 - Site differences in the fatty acid composition of subcutaneous adipose tissue of obese ...
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Archives of biochemistry and biophysics     Volume:  284     ISSN:  0003-9861     ISO Abbreviation:  Arch. Biochem. Biophys.     Publication Date:  1991 Jan 
Date Detail:
Created Date:  1991-02-28     Completed Date:  1991-02-28     Revised Date:  2000-12-18    
Medline Journal Info:
Nlm Unique ID:  0372430     Medline TA:  Arch Biochem Biophys     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  174-80     Citation Subset:  IM    
Laboratory of Biochemical Pharmacology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Circular Dichroism
DNA Mutational Analysis
Protein Conformation
Protein Denaturation
Salmonella typhimurium / enzymology*
Structure-Activity Relationship
Tryptophan Synthase / chemistry*
Reg. No./Substance:
EC Synthase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Previous Document:  Stereoisomerism in plant disease resistance: induction and isolation of the 7,2'-dihydroxy-4',5'-met...
Next Document:  Zeta-crystallin from guinea pig lens is capable of functioning catalytically as an oxidoreductase.