Visualizing a one-way protein encounter complex by ultrafast single-molecule mixing

被引：90

作者：

Gambin, Yann ^{[2
]}

VanDelinder, Virginia ^{[1
]}

Ferreon, Allan Chris M. ^{[2
]}

Lemke, Edward A. ^{[2
]}

Groisman, Alex ^{[1
]}

Deniz, Ashok A. ^{[2
]}

机构：

[1] Univ Calif San Diego, Dept Phys, San Diego, CA 92103 USA

[2] Scripps Res Inst, Dept Mol Biol, La Jolla, CA 92037 USA

来源：

NATURE METHODS | 2011年 / 8卷 / 03期

基金：

美国国家科学基金会; 美国国家卫生研究院;

关键词：

MICROFLUIDIC MIXER; FOLDING KINETICS; FLUORESCENCE; SPECTROSCOPY;

D O I：

10.1038/NMETH.1568

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We combined rapid microfluidic mixing with single-molecule fluorescence resonance energy transfer to study the folding kinetics of the intrinsically disordered human protein alpha-synuclein. The time-resolution of 0.2 ms revealed initial collapse of the unfolded protein induced by binding with lipid mimics and subsequent rapid formation of transient structures in the encounter complex. The method also enabled analysis of rapid dissociation and unfolding of weakly bound complexes triggered by massive dilution.

引用

页码：239 / U77

页数：6

共 15 条

[1] Experimental determination of upper bound for transition path times in protein folding from single-molecule photon-by-photon trajectories [J].

Chung, Hoi Sung ;

Louis, John M. ;

Eaton, William A. .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2009, 106 (29) :11837-11844

[2] Direct Observation of Downhill Folding of λ-Repressor in a Microfluidic Mixer [J].

DeCamp, Stephen J. ;

Naganathan, Athi N. ;

Waldauer, Steven A. ;

Bakajin, Olgica ;

Lapidus, Lisa J. .

BIOPHYSICAL JOURNAL, 2009, 97 (06) :1772-1777

[3] Single-molecule biophysics: at the interface of biology, physics and chemistry [J].

Deniz, Ashok A. ;

Mukhopadhyay, Samrat ;

Lemke, Edward A. .

JOURNAL OF THE ROYAL SOCIETY INTERFACE, 2008, 5 (18) :15-45

[4] α-Synuclein multistate folding thermodynamics:: Implications for protein misfolding and aggregation [J].

Ferreon, Allan Chris M. ;

Deniz, Ashok A. .

BIOCHEMISTRY, 2007, 46 (15) :4499-4509

[5] Interplay of α-synuclein binding and conformational switching probed by single-molecule fluorescence [J].

Ferreon, Allan Chris M. ;

Gambin, Yann ;

Lemke, Edward A. ;

Deniz, Ashok A. .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2009, 106 (14) :5645-5650

[6] Nonequilibrium single molecule protein folding in a coaxial mixer [J].

Hamadani, Kambiz M. ;

Weiss, Shimon .

BIOPHYSICAL JOURNAL, 2008, 95 (01) :352-365

[7] Optimization of a microfluidic mixer for studying protein folding kinetics [J].

Hertzog, David E. ;

Ivorra, Benjamin ;

Mohammadi, Bijan ;

Bakajin, Olgica ;

Santiago, Juan G. .

ANALYTICAL CHEMISTRY, 2006, 78 (13) :4299-4306

[8] Mapping protein collapse with single-molecule fluorescence and kinetic synchrotron radiation circular dichroism spectroscopy [J].

Hoffmann, Armin ;

Kane, Avinash ;

Nettels, Daniel ;

Hertzog, David E. ;

Baumgaertel, Peter ;

Lengefeld, Jan ;

Reichardt, Gerd ;

Horsley, David A. ;

Seckler, Robert ;

Bakajin, Olgica ;

Schuler, Benjamin .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2007, 104 (01) :105-110

[9] Single-molecule spectroscopy of protein folding in a chaperonin cage [J].

Hofmann, Hagen ;

Hillger, Frank ;

Pfeil, Shawn H. ;

Hoffmann, Armin ;

Streich, Daniel ;

Haenni, Dominik ;

Nettels, Daniel ;

Lipman, Everett A. ;

Schuler, Benjamin .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2010, 107 (26) :11793-11798

[10] Hydrodynamic focusing on a silicon chip: Mixing nanoliters in microseconds [J].

Knight, JB ;

Vishwanath, A ;

Brody, JP ;

Austin, RH .

PHYSICAL REVIEW LETTERS, 1998, 80 (17) :3863-3866

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