Thermal stability of dihydrofolate reductase and its fused proteins with oligopeptides.
Two fused proteins of dihydrofolate reductase (DHFR) with oligopeptides were prepared by a recombinant DNA method. One of these, DHFR-IQI, has three (Ile-Gln-Ile) and the other, DHFR-lek, has eight (Ile-Arg-Met-Tyr-Gly-Gly-Phe-Leu) additional amino acid residues at the C terminals; in both proteins, Cys152 of wild DHFR is replaced by Glu. The thermal transition of the proteins was measured by CD and DSC at pH 7.0 and compared with that of wild DHFR. The results show that the thermal stability of DHFR-IQI is the same as that of the wild DHFR and that of DHFR-lek is less than that of the former two DHFRs. Analysis of the DSC data of DHFR-IQI indicates that the thermal transition is a three-state one. Data from both DSC and CD measurements suggest the association of DHFR-lek molecules.
Uedaira
H
H
Research Institute for Polymers and Textiles, Ibaraki, Japan.
Kidokoro
S
S
Iwakura
M
M
Honda
S
S
Ohashi
S
S
D002152:Calorimetry, Differential Scanning; D002942:Circular Dichroism; D004795:Enzyme Stability; D008962:Models, Theoretical; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000378:metabolism; D013762:Tetrahydrofolate Dehydrogenase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013816:Thermodynamics
D011993:Recombinant Fusion Proteins; D013762:Tetrahydrofolate Dehydrogenase