Trimethylamine N-oxide-induced cooperative folding of an intrinsically unfolded transcription-activating fragment of human glucocorticoid receptor.
A number of biologically important proteins or protein domains identified recently are fully or partially unstructured (unfolded). Methods that allow studies of the propensity of such proteins to fold naturally are valuable. The traditional biophysical approaches using alcohols to drive alpha-helix formation raise serious questions of the relevance of alcohol-induced structure to the biologically important conformations. Recently we illustrated the extraordinary capability of the naturally occurring solute, trimethylamine N-oxide (TMAO), to force two unfolded proteins to fold to native-like species with significant functional activity. In the present work we apply this technique to recombinant human glucocorticoid receptor fragments consisting of residues 1-500 and residues 77-262. CD and fluorescence spectroscopy showed that both were largely disordered in aqueous solution. TMAO induced a condensed structure in the large fragment, indicated by the substantial enhancement in intrinsic fluorescence and blue shift of fluorescent maxima. CD spectroscopy demonstrated that the TMAO-induced structure is different from the alpha-helix-rich conformation driven by trifluoroethanol (TFE). In contrast to TFE, the conformational transition of the 1-500 fragment induced by TMAO is cooperative, a condition characteristic of proteins with unique structures.
Baskakov
I V
IV
Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston, Texas 77555-0645, USA.
Kumar
R
R
Srinivasan
G
G
Ji
Y S
YS
Bolen
D W
DW
Thompson
E B
EB
D002942:Circular Dichroism; D006801:Humans; D008744:Methylamines / Q000494:pharmacology*; D010446:Peptide Fragments / Q000378:metabolism; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011965:Receptors, Glucocorticoid / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism; D014158:Transcription, Genetic
D008744:Methylamines; D010446:Peptide Fragments; D011965:Receptors, Glucocorticoid; C005855:trimethyloxamine