Comparative Stability of Dihydrofolate Reductase Mutants in Vitro and in Vivo
Document Type
Article
Publication Date
1993
Digital Object Identifier (DOI)
https://doi.org/10.1093/protein/6.1.81
Abstract
Dihydrofolate reductase mutants with amino acid replacements in the active center (Thr35 ⇒ Asp mutant, Arg57 ⇒ His mutant and the mutant with triple replacement Thr35 ⇒ Asp, Asn37 ⇒ Ser, Arg57 ⇒ His) were obtained by site-directed mutagenesis. The stabilization effect of trimethoprim and NADP·H on the protein tertiary structure in vitro has been investigated. In the case of mutants with a ‘weak’ tertiary structure (Thr35 ⇒ Asp35 and the triple mutant) the separate addition of ligands does not affect their stability. The simultaneous addition of these ligands to Thr35 ⇒ Asp35 and the triple mutant leads to the large increase in their stability. A distinct correlation was found between the in vitro studied stability of the mutant proteins to the urea- or heat-induced denaturation and the level of proteolytic degradation of these mutants previously observed in vivo.
Was this content written or created while at USF?
Yes
Citation / Publisher Attribution
Protein Engineering, Design and Selection, v. 6, issue 1, p. 81-84
Scholar Commons Citation
Leontiev, Vladimir V.; Uversky, Vladimir N.; and Gudkov, Anatoly T., "Comparative Stability of Dihydrofolate Reductase Mutants in Vitro and in Vivo" (1993). Molecular Medicine Faculty Publications. 623.
https://digitalcommons.usf.edu/mme_facpub/623
