Abstract
Although often depicted as rigid structures, proteins are highly dynamic systems, whose motions are essential to their functions. Despite this, it is difficult to investigate protein dynamics due to the rapid timescale at which they sample their conformational space, leading most NMR-determined structures to represent only an averaged snapshot of the dynamic picture. While NMR relaxation measurements can help to determine local dynamics, it is difficult to detect translational or concerted motion, and only recently have significant advances been made to make it possible to acquire a more holistic representation of the dynamics and structural landscapes of proteins. Here, we briefly revisit our most recent progress in the theory and use of exact nuclear Overhauser enhancements (eNOEs) for the calculation of structural ensembles that describe their conformational space. New developments are primarily targeted at increasing the number and improving the quality of extracted eNOE distance restraints, such that the multi-state structure calculation can be applied to proteins of higher molecular weights. We then review the implications of the exact NOE to the protein dynamics and function of cyclophilin A and the WW domain of Pin1, and finally discuss our current research and future directions.
Highlights
Proteins exist in equilibrium between many structural states, and as such, are inherently dynamic systems
For nuclear Overhauser enhancement (NOE) with unresolved diagonals, we introduced a protocol to increase the number of exact nuclear Overhauser enhancements (eNOEs) that can be obtained from proteins of larger size, while avoiding adding semi-quantitative restraints, such as those obtained from conventional NOEs, that may induce distance errors into the structure
We have developed a method for stereospecific assignments for the majority of relevant diastereotopic groups by comparing eNOE-derived distances to protein structure bundles calculated without stereospecific assignments, making it possible to obtain more detailed structural and dynamical information from
Summary
Proteins exist in equilibrium between many structural states, and as such, are inherently dynamic systems. The calculated structures determined from conventional NOEs fail to represent the dynamic nature and exact structure of the proteins by which they are measured, and the protocol for the generation of atomic-resolution spatial representations still needs improvement, due to the difficulty in detecting translational and concerted motions. Using eNOEs for single-state structure calculations results in many distance restraint violations, which are indicative of the structure not agreeing well with the experimental data We found that these violations can be attributed to the motion-averaged nature of the measured eNOEs, which carry information about the spatial dynamics of mobile atoms in a protein. G (GB3; residues), the second post-synaptic density-95/discs calculated from eNOEs for the WW domain of Pin (34 residues), the third domain of immunoglobulin large/zonula occludens-1 (PDZ2) domain from human tyrosine phosphatase 1E (97 residues), and binding protein. Occludens-1 (PDZ2) domain from human tyrosine phosphatase 1E (97 residues), and human cyclophilin
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
