Dynamics of equilibrium folding and unfolding transitions of titin immunoglobulin domain under constant forces.
The mechanical stability of force-bearing proteins is crucial for their functions. However, slow transition rates of complex protein domains have made it challenging to investigate their equilibrium force-dependent structural transitions. Using ultra stable magnetic tweezers, we report the first equilibrium single-molecule force manipulation study of the classic titin I27 immunoglobulin domain. We found that individual I27 in a tandem repeat unfold/fold independently. We obtained the force-dependent free energy difference between unfolded and folded I27 and determined the critical force (∼5.4 pN) at which unfolding and folding have equal probability. We also determined the force-dependent free energy landscape of unfolding/folding transitions based on measurement of the free energy cost of unfolding. In addition to providing insights into the force-dependent structural transitions of titin I27, our results suggest that the conformations of titin immunoglobulin domains can be significantly altered during low force, long duration muscle stretching.
†Department of Physics, Xiamen University, Xiamen, Fujian 361005, China.|‡Mechanobiology Institute, National University of Singapore, Singapore 117411.
Hu
H
Chen
†Department of Physics, Xiamen University, Xiamen, Fujian 361005, China.|‡Mechanobiology Institute, National University of Singapore, Singapore 117411.
Guohua
G
Yuan
‡Mechanobiology Institute, National University of Singapore, Singapore 117411.
Ricksen S
RS
Winardhi
‡Mechanobiology Institute, National University of Singapore, Singapore 117411.
Mingxi
M
Yao
¶Department of Biological Sciences, Columbia University, New York, New York 10027, United States.
Ionel
I
Popa
¶Department of Biological Sciences, Columbia University, New York, New York 10027, United States.
Julio M
JM
Fernandez
‡Mechanobiology Institute, National University of Singapore, Singapore 117411.|§Department of Physics, National University of Singapore, Singapore 117542.|∥Centre for Bioimaging Sciences, National University of Singapore, Singapore 117546.
Jie
J
Yan
D001696:Biomechanical Phenomena; D064211:Connectin / Q000737:chemistry*; D004548:Elasticity; D007136:Immunoglobulins / Q000737:chemistry*; D055595:Mechanical Phenomena; D017434:Protein Structure, Tertiary; D058767:Protein Unfolding; D013816:Thermodynamics
D064211:Connectin; D007136:Immunoglobulins