Observation of protein folding/unfolding dynamics of ubiquitin trapped in agarose gel by single-molecule FRET

被引：8

作者：

Yang, Li-Ling ^{[1
,2
,3
,4
]}

Kao, Michael W. -P. ^{[1
]}

Chen, Hsin-Liang ^{[1
,2
]}

Lim, Tsong-Shin ^{[5
]}

Fann, Wunshain ^{[2
]}

Chen, Rita P. -Y. ^{[1
]}

机构：

[1] Acad Sinica, Inst Biol Chem, Taipei 115, Taiwan

[2] Acad Sinica, Inst Atom & Mol Sci, Taipei 106, Taiwan

[3] Acad Sinica, Taiwan Int Grad Program, Taipei 115, Taiwan

[4] Natl Taiwan Univ, Dept Phys, Taipei 106, Taiwan

[5] Tunghai Univ, Dept Phys, Taichung 407, Taiwan

来源：

EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS | 2012年 / 41卷 / 02期

关键词：

FRET; Ubiquitin; Folding; Single molecule; Conformational heterogeneity; Agarose; RESONANCE ENERGY-TRANSFER; ENZYME CONFORMATIONAL DYNAMICS; NUCLEAR MAGNETIC-RESONANCE; FLUORESCENCE SPECTROSCOPY; PORE-SIZE; SURFACE; DEPOLARIZATION; DENATURATION; FLUCTUATIONS; BIOMOLECULES;

D O I：

10.1007/s00249-011-0772-6

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

A ubiquitin mutant with two Cys mutations, m[C]q/S65C, was site-specifically labeled with two dye molecules, Alexa Fluor 488 (donor) and Alexa Fluor 594 (acceptor), due to the different reactivity of these two Cys residues. This doubly dye-labeled ubiquitin has lower structural stability than wild-type ubiquitin. Taking advantage of this decreased stability, conformational heterogeneity of this protein under nondenaturing condition was observed at the single-molecule level using single-paired Forster resonance energy transfer (FRET) by trapping the protein in agarose gel. Three conformational populations corresponding to folded (E (ET) a parts per thousand 0.95), loosely packed (E (ET) a parts per thousand 0.72), and unfolded (E (ET) a parts per thousand 0.22) structures, and the structural transitions between them were observed. Our results suggest that agarose immobilization is good for observing structural dynamics of proteins under native condition.

引用

页码：189 / 198

页数：10

共 50 条

[1] Observation of protein folding/unfolding dynamics of ubiquitin trapped in agarose gel by single-molecule FRET
Li-Ling Yang
Michael W.-P. Kao
Hsin-Liang Chen
Tsong-Shin Lim
Wunshain Fann
Rita P.-Y. Chen
European Biophysics Journal, 2012, 41 : 189 - 198
[2] SINGLE-MOLECULE FRET OF PROTEIN-FOLDING DYNAMICS
Nettels, Daniel
Schuler, Benjamin
SINGLE-MOLECULE BIOPHYSICS: EXPERIMENT AND THEORY, VOL 146, 2012, 146 : 23 - 48
[3] Unfolding/Folding of a Multi-Domain Protein Untangled by Single-Molecule FRET
Schoene, Antonie
Kempe, Daryan
Cerminara, Michele
Gabba, Matteo
Zuechner, Tina
Fitter, Joerg
BIOPHYSICAL JOURNAL, 2016, 110 (03) : 636A - 636A
[4] Protein folding studied by single-molecule FRET
Schuler, Benjamin
Eaton, William A.
CURRENT OPINION IN STRUCTURAL BIOLOGY, 2008, 18 (01) : 16 - 26
[5] Single Molecule FRET of Protein Folding Dynamics
Schuler, Ben
FASEB JOURNAL, 2009, 23
[6] Single-molecule FRET and transition paths in protein folding
Eaton, W. A.
FEBS JOURNAL, 2011, 278 : 13 - 13
[7] Single-molecule FRET and transition paths in protein folding
Chung, Hoi Sung
Eaton, William A.
ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 2011, 242
[8] Structural dynamics of membrane-protein folding from single-molecule FRET
Krainer, Georg
Keller, Sandro
Schlierf, Michael
CURRENT OPINION IN STRUCTURAL BIOLOGY, 2019, 58 : 124 - 137
[9] Multicolor single-molecule FRET to explore protein folding and binding
Gambin, Yann
Deniz, Ashok A.
MOLECULAR BIOSYSTEMS, 2010, 6 (09) : 1540 - 1547
[10] Mechanical folding and unfolding of protein barnase at the single-molecule level
Alemany, A.
Rey, B.
Frutos, S.
Cecconi, C.
Ritort, F.
EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS, 2015, 44 : S229 - S229

← 1 2 3 4 5 →