Does trifluoroethanol affect folding pathways and can it be used as a probe of structure in transition states?


Abstract

Nonaqueous co-solvents, particularly 2,2,2-trifluoroethanol (TFE), have been used as tools to study protein folding. By analyzing FKBP12, an alpha/beta-protein that folds with two-state kinetics, we have been able to address three key questions concerning the use of TFE. First, does TFE perturb the folding pathway? Second, can the observed changes in the rate of folding and unfolding in TFE be attributed to a change in free energy of a single state? Finally, can TFE be used to infer information on secondary structure formation in the transition state? Protein engineering experiments on FKBP12, coupled with folding and unfolding experiments in 0% and 9.6% TFE, conclusively show that TFE does not perturb the folding pathway of this protein. Our results also suggest that the changes in folding and unfolding rates observed in 9.6% TFE are due to a global effect of TFE on the protein, rather than the stabilization of any elements of secondary structure in the transition state. Thus, studies with TFE and other co-solvents can be accurately interpreted only when combined with other techniques. Study holds ProTherm entries: 15054, 15055, 15056, 15057, 15058, 15059, 15060, 15061, 15062, 15063, 15064, 15065, 15066, 15067, 15068, 15069, 15070, 15071, 15072, 15073, 15074, 15075, 15076, 15077, 15078, 15079, 15080, 15081 Extra Details: dithiothreitol (DTT)(1 mM) was added in the experiment,Calculated in 0% TFE using an average mU-F (1.59 } 0.02) trifluoroethanol, protein folding, FKBP12, transition state, secondary structure

Submission Details

ID: zDRNuTyv3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:45 p.m.

Version: 1

Publication Details
Main ER;Jackson SE,Nat. Struct. Biol. (1999) Does trifluoroethanol affect folding pathways and can it be used as a probe of structure in transition states? PMID:10467094
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
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
1FKL 1995-08-18T00:00:00+0000 1.7 ATOMIC STRUCTURE OF FKBP12-RAPAYMYCIN, AN IMMUNOPHILIN-IMMUNOSUPPRESSANT COMPLEX
1TCO 1996-08-21T00:00:00+0000 2.5 TERNARY COMPLEX OF A CALCINEURIN A FRAGMENT, CALCINEURIN B, FKBP12 AND THE IMMUNOSUPPRESSANT DRUG FK506 (TACROLIMUS)
1FKK 1995-08-18T00:00:00+0000 2.2 ATOMIC STRUCTURE OF FKBP12, AN IMMUNOPHILIN BINDING PROTEIN
3J8H 2014-10-26T00:00:00+0000 3.8 Structure of the rabbit ryanodine receptor RyR1 in complex with FKBP12 at 3.8 Angstrom resolution
5GKY 2016-07-07T00:00:00+0000 3.8 Structure of RyR1 in a closed state (C1 conformer)
5GKZ 2016-07-07T00:00:00+0000 4.0 Structure of RyR1 in a closed state (C3 conformer)
5GL1 2016-07-07T00:00:00+0000 5.7 Structure of RyR1 in an open state
5GL0 2016-07-07T00:00:00+0000 4.2 Structure of RyR1 in a closed state (C4 conformer)
2FKE 1993-01-27T00:00:00+0000 1.72 FK-506-BINDING PROTEIN: THREE-DIMENSIONAL STRUCTURE OF THE COMPLEX WITH THE ANTAGONIST L-685,818
1J4H 2001-09-30T00:00:00+0000 1.8 crystal structure analysis of the FKBP12 complexed with 000107 small molecule

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
97.2 Peptidyl-prolyl cis-trans isomerase FKBP1A Q62658 FKB1A_RAT
97.2 Peptidyl-prolyl cis-trans isomerase FKBP1A P26883 FKB1A_MOUSE
97.2 Peptidyl-prolyl cis-trans isomerase FKBP1A P18203 FKB1A_BOVIN
100.0 Peptidyl-prolyl cis-trans isomerase FKBP1A P62943 FKB1A_RABIT
100.0 Peptidyl-prolyl cis-trans isomerase FKBP1A P62942 FKB1A_HUMAN