A hydrophobic cluster forms early in the folding of dihydrofolate reductase.


Abstract

The rapid kinetic phase that leads from unfolded species to transient folding intermediates in dihydrofolate reductase from Escherichia coli was examined by site-directed mutagenesis and by physicochemical means. The absence of this fluorescence-detected phase in the refolding of the Trp-74Phe mutant protein strongly implies that this early phase in refolding can be assigned to just one of the five Trp residues in the protein, Trp-74. In addition, water-soluble fluorescence quenching agents, iodide and cesium, have a much less significant effect on this early step in refolding than on the slower phases that lead to native and native-like conformers. These and other data imply that an important early event in the folding of dihydrofolate reductase is the formation of a hydrophobic cluster which protects Trp-74 from solvent. Study holds ProTherm entries: 10792, 10793 Extra Details: additive : 2-mercaptethanol(1 mM), protein folding; folding intermediates; hydrophobic effect;,tryptophan fluorescence; site-directed mutagenesis

Submission Details

ID: 6gpxRwNz

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:40 p.m.

Version: 1

Publication Details
Garvey EP;Swank J;Matthews CR,Proteins (1989) A hydrophobic cluster forms early in the folding of dihydrofolate reductase. PMID:2622906
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