Complex bio-macromolecules such as membrane proteins play crucial roles in many cellular and physiological processes and specific defects are associated with many known. Determining their three- dimensional structures is one of the main objectives in structural biology and the NIH devotes considerable resources towards this goal. In recent years, pulse EPR spectroscopy, in particular Double Electron-Electron Resonance (DEER) has contributed valuable structural constraints to solve structures of bio-macromolecules. However, currently these methods are only available to experts in the field because operating the complex instrument requires lots of training making it difficult for a non-EPR expert to access this highly valuable information. State-of-the-art spectrometer configurations may be the most versatile option, however, many features of current commercially available systems are unnecessary or overly complicated. Options such as continuous wave (cw) EPR are unnecessary, when the main objective is to measure distances. In addition, it is also often overwhelming to applications scientist that are not experts in pulsed EPR spectroscopy. Today, much of the experiment setup and processing can be automated and standardized. Current commercial solutions do not come equipped with these streamlined features. With DEER becoming more popular it is time to propose a solution that is geared specifically towards distance measurements than creating a universal research instruments. Removing unnecessary options and focusing only on DEER spectroscopy will allow us to design a compact, turn-key instrument. In this SBIR application we propose to develop a compact, turn-key Q-band pulsed EPR spectrometer for pulsed dipolar spectroscopy. The system will feature a liquid cryogen-free magnet and cryostat, a low-Q but high-sensitivity resonator and a compact, EPR bridge with state-of-the-art pulse shaping capabilities. The system will fully computer controlled and easy to operate with minimal training. The successful development of this compact, Q-band pulsed EPR spectrometer will provide researchers access to turn-key instrumentation allowing them to easily incorporate experiments such as DEER in their research. This will greatly proliferate the method and is of large interest to many projects funded by the U.S. National Institutes of Health.
Public Health Relevance Statement: Project Summary / Abstract Pulsed Dipolar Spectroscopy such as Double Electron-Electron Resonance (DEER) is a valuable technique to obtain structural constraints of complex bio-macromolecules. While the technique is versatile and powerful the field lacks simple, turn-key instrumentation dedicated to this technique. This severely slows down the proliferation of the technique. The proposed instrument is a cost-effective solution to this problem. It will make the method available to a much broader audience and proliferate research that is at the heart of the National Institutes of Health.
Project Terms: Alzheimer's Disease; AD dementia; Alzheimer; Alzheimer Type Dementia; Alzheimer disease; Alzheimer sclerosis; Alzheimer syndrome; Alzheimer's; Alzheimer's disease dementia; Alzheimers Dementia; Alzheimers disease; Primary Senile Degenerative Dementia; dementia of the Alzheimer type; primary degenerative dementia; senile dementia of the Alzheimer type; Cell physiology; Cell Function; Cell Process; Cellular Function; Cellular Physiology; Cellular Process; Subcellular Process; Computers; Non-Insulin-Dependent Diabetes Mellitus; Adult-Onset Diabetes Mellitus; Ketosis-Resistant Diabetes Mellitus; Maturity-Onset Diabetes Mellitus; NIDDM; Non-Insulin Dependent Diabetes; Noninsulin Dependent Diabetes; Noninsulin Dependent Diabetes Mellitus; Slow-Onset Diabetes Mellitus; Stable Diabetes Mellitus; T2 DM; T2D; T2DM; Type 2 Diabetes Mellitus; Type 2 diabetes; Type II Diabetes Mellitus; Type II diabetes; adult onset diabetes; ketosis resistant diabetes; maturity onset diabetes; type 2 DM; type II DM; type two diabetes; Disease; Disorder; Electromagnetics; Electron Spin Resonance Spectroscopy; EPR spectroscopy; ESR Spectroscopy; Electron Paramagnetic Resonance; Electron Spin Resonance; Paramagnetic Resonance; electron paramagnetic resonance spectroscopy; Electrons; Negative Beta Particle; Negatrons; Elements; Extravasation; Leakage; Spillage; Goals; Hand; Health; Heart; instrumentation; Magnetic Resonance Spectroscopy; MR Spectroscopy; Membrane Proteins; Membrane Protein Gene; Membrane-Associated Proteins; Surface Proteins; Methods; microwave electromagnetic radiation; Microwave Electromagnetic; Microwaves; microwave radiation; United States National Institutes of Health; NIH; National Institutes of Health; Parkinson Disease; Paralysis Agitans; Parkinson; Parkinson's disease; Parkinsons disease; Primary Parkinsonism; Physiological Processes; Organism-Level Process; Organismal Process; Physiologic Processes; Play; Problem Solving; Proteins; Research; Research Personnel; Investigators; Researchers; Resources; Research Resources; Role; social role; Signal Transduction; Cell Communication and Signaling; Cell Signaling; Intracellular Communication and Signaling; Signal Transduction Systems; Signaling; biological signal transduction; Spectrum Analysis; Spectroscopy; Spectrum Analyses; Spin Labels; Standardization; Mechanical Stress; Temperature; Time; Measures; Morphologic artifacts; Artifacts; base; macromolecule; cryostat; improved; Site; Residual; Residual state; Phase; Variation; Variant; Single Crystal Diffraction; X Ray Crystallographies; X-Ray Diffraction Crystallography; X-Ray/Neutron Crystallography; Xray Crystallography; X-Ray Crystallography; Training; Measurement; Funding; Collaborations; fluid; liquid; Liquid substance; Shapes; tool; electric field; instrument; programs; mechanical; Mechanics; Pulse; Physiologic pulse; Scientist; Frequencies; Complex; Techniques; System; interest; Performance; structural biology; Cryo-electron Microscopy; Electron Cryomicroscopy; cryo-EM; cryoEM; Cryoelectron Microscopy; Structure; simulation; Sampling; NMR Spectrometer; nuclear magnetic resonance spectroscopy; NMR Spectroscopy; preventing; prevent; Defect; protein structures; proteins structure; protein structure; Proliferating; Protein Dynamics; Resolution; SBIR; Small Business Innovation Research; Small Business Innovation Research Grant; developmental; Development; vector; designing; design; 3-D structure; 3-dimensional structure; 3D structure; three dimensional structure; cost effective; Coupling; prototype; operation; experiment; experimental research; experimental study
