The effect of charge-introduction mutations on E. coli thioredoxin stability.
Technological applications of proteins are often hampered by their low-stability and, consequently, the development of procedures for protein stabilization is of considerable biotechnological interest. Here, we use simple electrostatics to determine positions in E. coli thioredoxin at which mutations that introduce new charged residues are expected to lead to stability enhancement. We also obtain the corresponding mutants and characterize their stability using differential scanning calorimetry. The results are interpreted in terms of the accessibility in the native structure of the mutated residues and the potential effect of the mutations on the residual structure of the denatured state.
Perez-Jimenez
Raul
R
Facultad de Ciencias, Departamento de Quimica Fisica, 18071-Granada, Spain.
Godoy-Ruiz
Raquel
R
Ibarra-Molero
Beatriz
B
Sanchez-Ruiz
Jose M
JM
D002152:Calorimetry, Differential Scanning; D004926:Escherichia coli / Q000235:genetics*; D008239:Lysine / Q000235:genetics / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D011489:Protein Denaturation; D015202:Protein Engineering; D055672:Static Electricity; D013816:Thermodynamics; D013879:Thioredoxins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism
D013879:Thioredoxins; D008239:Lysine