dihydrofolate reductase hvDHFR1 and hvDHFR2 unfold at KCl concentrations below 0.5 M. Above 1 M, the KCl dependence of the dihydrofolate reductase activities can be attributed to the effect of salt on substrate affinity
dihydrofolate reductase hvDHFR1 is ,12 kcal/mol more stable to urea denaturation than dihydrofolate reductase hvDHFR2. NaCl, KCl, and CsCl enhance the stability to urea denaturation
large quantities of recombinant wild-type enzyme from Escherichia coli, purification of mutants L30K, A31K and double mutant L30K/A31K recombinant from Escherichia coli
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RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
study on kinetic folding of urea-denaturedc dihydrofolate reductase and comparison with Escherichia coli enzyme. Folding follows similar kinetics for both enzymes, with a 5-ms stopped-flow burst-phase species that folds to the native state through two sequential intermediateswith relaxation times of 0.1-3 sec and 25-100 sec. The unfolding of Haloferax volcanii enzyme at low ionic strength is relatively slow. Increased KCl concentrations slow the urea-induced unfolding of both enzymes, but much less than expected from equilibrium studies. Unfolding rates are relatively independent of ionic strength
very fast and total reconstitution of active recombinant wild-type enzyme from inclusion bodies due to overexpression in E. coli by 6 M guanidine hydrochloride followed by dilution into 1 M NaCl or KCl solution
High expression in Escherichia coli of the gene coding for dihydrofolate reductase of the extremely halophilic archaebacterium Haloferax volcanii. Reconstitution of the active enzyme and mutation studies
Kinetic folding of Haloferax volcanii and Escherichia coli dihydrofolate reductases: haloadaptation by unfolded state destabilization at high ionic strength
Pieper, U.; Kapadia, G.; Mevarech, M.; Herzberg, O.
Structural features of halophilicity derived from the crystal structure of dihydrofolate reductase from the Dead Sea halophilic archaeon, Haloferax volcanii