Reversible dissociation and unfolding of the dimeric protein thymidylate synthase
Open Access
- 1 June 1992
- journal article
- research article
- Published by Wiley in Protein Science
- Vol. 1 (6), 796-800
- https://doi.org/10.1002/pro.5560010611
Abstract
Conditions for in vitro unfolding and refolding of dimeric thymidylate synthase from Lactobacillus casei were found. Ultraviolet difference and circular dichroism spectra showed that the enzyme was completely unfolded at concentrations of urea over 5.5 M. As measured by restoration of enzyme activity, refolding was accomplished when 0.5 M potassium chloride was included in the refolding mixture. Recombination of subunits from catalytically inactive mutant homodimers to form an active hybrid dimer was achieved under these unfolding–refolding conditions, demonstrating a monomer to dimer association step.Keywords
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