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:2622906Additional Information
Study Summary
| Number of data points | 3 |
| Proteins | Dihydrofolate reductase ; Dihydrofolate reductase |
| Unique complexes | 2 |
| Assays/Quantities/Protocols | Experimental Assay: ddG_H2O ; Experimental Assay: Cm |
| Libraries | Mutations for sequence MISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLDKPVIMGRHTWESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLPKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR |
Structure view and single mutant data analysis
Study data
| Colors: | D | E | R | H | K | S | T | N | Q | A | V | I | L | M | F | Y | W | C | G | P |
|---|
Data Distribution
Studies with similar sequences (approximate matches)
Correlation with other assays (exact sequence matches)
Relevant PDB Entries
| Structure ID | Release Date | Resolution | Structure Title |
|---|---|---|---|
| 6CW7 | 2018-03-30T00:00:00+0000 | 1.03 | E. coli DHFR product complex with (6S)-5,6,7,8-TETRAHYDROFOLATE |
| 6CXK | 2018-04-03T00:00:00+0000 | 1.11 | E. coli DHFR substrate complex with Dihydrofolate |
| 6CYV | 2018-04-06T00:00:00+0000 | 1.3 | E. coli DHFR ternary complex with NADP and dihydrofolate |
| 1DDR | 1995-06-29T00:00:00+0000 | 2.45 | MOLECULE: DIHYDROFOLATE REDUCTASE (E.C.1.5.1.3) COMPLEXED WITH METHOTREXATE AND UREA |
| 1DDS | 1995-06-29T00:00:00+0000 | 2.2 | MOLECULE: DIHYDROFOLATE REDUCTASE (E.C.1.5.1.3) COMPLEXED WITH METHOTREXATE |
| 1DHI | 1993-10-29T00:00:00+0000 | 1.9 | LONG-RANGE STRUCTURAL EFFECTS IN A SECOND-SITE REVERTANT OF A MUTANT DIHYDROFOLATE REDUCTASE |
| 1DHJ | 1993-10-29T00:00:00+0000 | 1.8 | LONG-RANGE STRUCTURAL EFFECTS IN A SECOND-SITE REVERTANT OF A MUTANT DIHYDROFOLATE REDUCTASE |
| 1DRA | 1991-11-06T00:00:00+0000 | 1.9 | CRYSTAL STRUCTURE OF UNLIGANDED ESCHERICHIA COLI DIHYDROFOLATE REDUCTASE. LIGAND-INDUCED CONFORMATIONAL CHANGES AND COOPERATIVITY IN BINDING |
| 1DRB | 1991-11-06T00:00:00+0000 | 1.96 | CRYSTAL STRUCTURE OF UNLIGANDED ESCHERICHIA COLI DIHYDROFOLATE REDUCTASE. LIGAND-INDUCED CONFORMATIONAL CHANGES AND COOPERATIVITY IN BINDING |
| 1DRE | 1996-11-28T00:00:00+0000 | 2.6 | DIHYDROFOLATE REDUCTASE COMPLEXED WITH METHOTREXATE AND NICOTINAMIDE ADENINE DINUCLEOTIDE PHOSPHATE (OXIDIZED FORM) |