The effects of macromolecular crowding on the mechanical stability of protein molecules.
Macromolecular crowding, a common phenomenon in the cellular environments, can significantly affect the thermodynamic and kinetic properties of proteins. A single-molecule method based on atomic force microscopy (AFM) was used to investigate the effects of macromolecular crowding on the forces required to unfold individual protein molecules. It was found that the mechanical stability of ubiquitin molecules was enhanced by macromolecular crowding from added dextran molecules. The average unfolding force increased from 210 pN in the absence of dextran to 234 pN in the presence of 300 g/L dextran at a pulling speed of 0.25 microm/sec. A theoretical model, accounting for the effects of macromolecular crowding on the native and transition states of the protein molecule by applying the scaled-particle theory, was used to quantitatively explain the crowding-induced increase in the unfolding force. The experimental results and interpretation presented could have wide implications for the many proteins that experience mechanical stresses and perform mechanical functions in the crowded environment of the cell.
Yuan
Jian-Min
JM
Department of Physics, Drexel University, Philadelphia, Pennsylvania 19104, USA.
Chyan
Chia-Lin
CL
Zhou
Huan-Xiang
HX
Chung
Tse-Yu
TY
Peng
Haibo
H
Ping
Guanghui
G
Yang
Guoliang
G
D002942:Circular Dichroism; D003911:Dextrans / Q000737:chemistry; D007700:Kinetics; D018625:Microscopy, Atomic Force; D020527:Microscopy, Scanning Probe; D017510:Protein Folding; D055550:Protein Stability; D017433:Protein Structure, Secondary; D013816:Thermodynamics; D025801:Ubiquitin / Q000737:chemistry*
D003911:Dextrans; D025801:Ubiquitin
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