Title
Slow Motion Protein Dance Visualized Using Red-Edge Excitation Shift of a Buried Fluorophore
Author
Prajna Mishra, Santosh Kumar Jha
Year
2019
Journal
The Journal of Physical Chemistry B
Abstract
It has been extremely challenging to detect protein structures with a dynamic core, such as dry molten globules, that remain in equilibrium with the tightly packed native (N) state and that are important for a myriad of entropy-driven protein functions. Here, we detect the higher entropy conformations of a human serum protein, using red-edge excitation shift experiments. We covalently introduced a fluorophore inside the protein core and observed that in a subset of native population, the side chains of the polar and buried residues have different spatial arrangements than the mean population and that they solvate the fluorophore on a timescale much slower than the nanosecond timescale of fluorescence. Our results provide direct evidence for the dense fluidity of protein core and show that alternate side-chain packing arrangements exist in the core that might be important for multiple binding functions of this protein.
Instrument
J-815
Keywords
Circular dichroism, Secondary structure, Chemical stability, Biochemistry