D-DNP for protein NMR
Nuclear magnetic resonance (NMR) spectroscopy presently stands as the sole high-resolution technique for deciphering the structural dynamics of proteins and nucleic acids in solution. Our strategy involves the implementation of dissolution dynamic nuclear polarization (D-DNP) to enhance NMR investigations of proteins. This methodology has been coined as HYPEX (hyperpolarization exchange spectroscopy), wherein a hyperpolarized aqueous buffer serves to amplify NMR signals by multiple orders of magnitude. The utilization of HYPEX NMR facilitates time-resolved examinations of macromolecules within conditions that mimic physiological settings, thereby enabling the precise determination of interaction kinetics.
Biomineralization and Biomimetic Materials
Due to its adherence to non-classical crystallization pathways, the process of biomineral nucleation, wherein soluble prenucleation inorganic entities undergo evolution into mineral phases under the precise governance of proteins, remains a subject of intense debate. The primary objective of this project is to elucidate the intricate mechanisms through which mineralizing proteins intricately control the configuration and dynamics of prenucleation inorganic entities during the initial phases of apatite development. This scrutiny will span from the solution phase to the point of nucleation, where the transition to the solid phase occurs.
The unveiling of these mechanisms is poised to illuminate the crucial early stages of apatite formation with an unprecedented level of detail in both spatial and temporal dimensions. Furthermore, this exploration holds the potential to enhance our understanding of bone-related disorders that arise due to aberrant mineral nucleation events.
Integrative Magnetic Resonance
While Nuclear Magnetic Resonance (NMR) spectroscopy detects nuclei within biomolecules, Electron Paramagnetic Resonance (EPR) spectroscopy exhibits sensitivity to distinct paramagnetic protein markers. EPR and NMR, as complementary magnetic resonance methodologies, operate across different time and length scales, collectively encompassing a broad spectrum of distances spanning from 0.1 Å to 10 nm. Moreover, they capture molecular motions ranging from picoseconds to seconds.
Our efforts are focused on pioneering applications that integrate both EPR and NMR techniques in the realm of protein structural biology. This innovative approach affords profound insights into the dynamic structures of proteins, encompassing both well-structured and intrinsically disordered proteins (IDPs). Through this concerted utilization of EPR and NMR, we unlock a comprehensive understanding of protein behaviors and conformations, fostering advancements in our comprehension of biomolecular systems.