Circularly Permuted Dihydrofolate Reductase of e.coli Has Functional Activity and a Destabilized Tertiary Structure
Document Type
Article
Publication Date
1994
Digital Object Identifier (DOI)
https://doi.org/10.1093/protein/7.11.1373
Abstract
Three circularly permuted variants of Escherichia coli dihydrofolate reductase genes were constructed. Linkers coding tri- and pentapeptides were used to connect the natural 5'- and 3'-terminal ends. Only one variant of circularly permuted protein with tripeptide linker and the cleavage of the peptide bond between 107 and 108 amino acid residues was produced in a good yield. The expressed protein was insoluble in the cells, but at pH 8.0 and higher the isolated protein was soluble. Enzymatic assay and physical studies have shown that permuted dihydrofolate reductase has a destabilized tertiary structure. Only the addition of the natural substrates or Inhibitors lead to the protein with the native-like structure and functional activity.
Was this content written or created while at USF?
Yes
Citation / Publisher Attribution
Protein Engineering, Design and Selection, v. 7, issue 11, p. 1373-1377
Scholar Commons Citation
Protasova, Natalya YU.; Kireeva, M. L.; Murzina, N. V.; Murzin, N. V.; Uversky, Vladimir N.; Gryaznova, O. I.; and Gudkov, Anatoly T., "Circularly Permuted Dihydrofolate Reductase of e.coli Has Functional Activity and a Destabilized Tertiary Structure" (1994). Molecular Medicine Faculty Publications. 627.
https://digitalcommons.usf.edu/mme_facpub/627
