GSBS Student Publications
Title
Microsecond acquisition of heterogeneous structure in the folding of a TIM barrel protein
Publication Date
2008-09-02
UMMS Affiliation
Graduate School of Biomedical Sciences; Department of Biochemistry and Molecular Pharmacology
Document Type
Article
Disciplines
Life Sciences | Medicine and Health Sciences
Abstract
The earliest kinetic folding events for (betaalpha)(8) barrels reflect the appearance of off-pathway intermediates. Continuous-flow microchannel mixing methods interfaced to small-angle x-ray scattering (SAXS), circular dichroism (CD), time-resolved Forster resonant energy transfer (trFRET), and time-resolved fluorescence anisotropy (trFLAN) have been used to directly monitor global and specific dimensional properties of the partially folded state in the microsecond time range for a representative (betaalpha)(8) barrel protein. Within 150 micros, the alpha-subunit of Trp synthase (alphaTS) experiences a global collapse and the partial formation of secondary structure. The time resolution of the folding reaction was enhanced with trFRET and trFLAN to show that, within 30 micros, a distinct and autonomous partially collapsed structure has already formed in the N-terminal and central regions but not in the C-terminal region. A distance distribution analysis of the trFRET data confirmed the presence of a heterogeneous ensemble that persists for several hundreds of microseconds. Ready access to locally folded, stable substructures may be a hallmark of repeat-module proteins and the source of early kinetic traps in these very common motifs. Their folding free-energy landscapes should be elaborated to capture this source of frustration.
DOI of Published Version
10.1073/pnas.0802788105
Source
Proc Natl Acad Sci U S A. 2008 Sep 9;105(36):13367-72. Epub 2008 Aug 29. Link to article on publisher's site
Journal/Book/Conference Title
Proceedings of the National Academy of Sciences of the United States of America
Related Resources
PubMed ID
18757725
Repository Citation
Wu, Ying; Kondrashkina, Elena; Kayatekin, Can; Matthews, C. Robert; and Bilsel, Osman, "Microsecond acquisition of heterogeneous structure in the folding of a TIM barrel protein" (2008). GSBS Student Publications. 1385.
https://escholarship.umassmed.edu/gsbs_sp/1385