Benchtop Q-Band Pulsed EPR Spectrometer for Intermolecular Distance Measurements

Project Summary / AbstractComplex 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.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.