Full distance-resolved folding energy landscape of one single protein molecule

被引：163

作者：

Gebhardt, J. Christof M. ^{[1
,2
]}

Bornschloegla, Thomas ^{[1
]}

Rief, Matthias ^{[1
]}

机构：

[1] Tech Univ Munich, Phys Dept E22, D-85748 Garching, Germany

[2] Munich Ctr Integrated Prot Sci, D-81377 Munich, Germany

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2010年 / 107卷 / 05期

关键词：

leucine zipper; force spectroscopy; optical tweezers; protein folding; deconvolution; GCN4; LEUCINE-ZIPPER; COILED-COIL PEPTIDE; FORCE SPECTROSCOPY; FLUCTUATION THEOREM; KINETICS; BARRIER; SURFACE; TITIN;

D O I：

10.1073/pnas.0909854107

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Kinetic bulk and single molecule folding experiments characterize barrier properties but the shape of folding landscapes between barrier top and native state is difficult to access. Here, we directly extract the full free energy landscape of a single molecule of the GCN4 leucine zipper using dual beam optical tweezers. To this end, we use deconvolution force spectroscopy to follow an individual molecule's trajectory with high temporal and spatial resolution. We find a heterogeneous energy landscape of the GCN4 leucine zipper domain. The energy profile is divided into two stable C-terminal heptad repeats and two less stable repeats at the N-terminus. Energies and transition barrier positions were confirmed by single molecule kinetic analysis. We anticipate that deconvolution sampling is a powerful tool for the model-free investigation of protein energy landscapes.

引用

页码：2013 / 2018

页数：6

共 42 条

[21] Ligand-Dependent Equilibrium Fluctuations of Single Calmodulin Molecules [J].

Junker, Jan Philipp ;

Ziegler, Fabian ;

Rief, Matthias .

SCIENCE, 2009, 323 (5914) :633-637

[22] An autonomous folding unit mediates the assembly of two-stranded coiled coils [J].

Kammerer, RA ;

Schulthess, T ;

Landwehr, R ;

Lustig, A ;

Engel, J ;

Aebi, U ;

Steinmetz, MO .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (23) :13419-13424

[23] A buried polar residue in the hydrophobic interface of the coiled-coil peptide, GCN4-p1, plays a thermodynamic, not a kinetic role in folding [J].

Knappenberger, JA ;

Smith, JE ;

Thorpe, SH ;

Zitzewitz, JA ;

Matthews, CR .

JOURNAL OF MOLECULAR BIOLOGY, 2002, 321 (01) :1-6

[24]

LEE DL, J MOL BIOL, V306, P539

[25] Barrier-limited, microsecond folding of a stable protein measured with hydrogen exchange: Implications for downhill folding [J].

Meisner, WK ;

Sosnick, TR .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (44) :15639-15644

[26] Energy landscapes of receptor-ligand bonds explored with dynamic force spectroscopy [J].

Merkel, R ;

Nassoy, P ;

Leung, A ;

Ritchie, K ;

Evans, E .

NATURE, 1999, 397 (6714) :50-53

[27] Differential detection of dual traps improves the spatial resolution of optical tweezers [J].

Moffitt, Jeffrey R. ;

Chemla, Yann R. ;

Izhaky, David ;

Bustamante, Carlos .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2006, 103 (24) :9006-9011

[28] Transition state heterogeneity in GCN4 coiled coil folding studied by using multisite mutations and crosslinking [J].

Moran, LB ;

Schneider, JP ;

Kentsis, A ;

Reddy, GA ;

Sosnick, TR .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (19) :10699-10704

[29] Unfolding pathways of individual bacteriorhodopsins [J].

Oesterhelt, F ;

Oesterhelt, D ;

Pfeiffer, M ;

Engel, A ;

Gaub, HE ;

Müller, DJ .

SCIENCE, 2000, 288 (5463) :143-146

[30] Theory of protein folding: The energy landscape perspective [J].

Onuchic, JN ;

LutheySchulten, Z ;

Wolynes, PG .

ANNUAL REVIEW OF PHYSICAL CHEMISTRY, 1997, 48 :545-600

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