Beckman Optima Multiwavelength Analytical Ultracentrifuge

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Project Summary/Abstract At the well-established X-ray crystallography facility at The Pennsylvania State University, we seek funds for a Beckman Coulter multiwavelength Optima analytical ultracentrifuge (AUC) instrument equipped with both UV-visible absorbance and Rayleigh interference optics. Partnered with the robust AUC software, UltraScan and SedFIT, this will enable accurate size, shape, partial specific volume, conformational change, molar mass, heterogeneity, binding thermodynamics and stoichiometry data on a host of disease related biomolecules, bacterial, viral, motor, nucleosomal and intrinsically disordered proteins, viral capsids, nucleic acids, liposomes and their complexes. No other functional AUC equipment exists on any of our twenty-four Penn State campuses. A modern AUC capability, will enable us to cater competently to the biophysical and structural characterization needs of our many exceptional NIH funded faculty and support them in an integrated structural biology research. We have identified the Beckman AUC Optima as the sole commercial product that meets our requirements, is state-of-the-art and cost-effective. It features 0.001 cm radial resolution, scanning speeds up to 8 sec/channel in absorbance mode, and 5 sec/channel in interference mode. The high scan-speed, good resolution and superior sensitivity is paramount for the success of the twenty-two cutting edge health care NIH projects and their applications amenable to multiwavelength extinction detection including (a) protein complexes and oligomers in the Dokholyan, Hancock, Weinert, Boehr, Cotruvo, Zhang and Nixon labs (b) protein nucleic acid complexes in the Tan, Krasilnikov, Murakami, Grigoryev and Moustafa labs (c) liquid-liquid phase separated systems in the Bevilacqua lab (d) intrinsically disordered proteins in the Showalter and Zhang labs (e) liposomes in the Tian lab and (f) anaerobic metalloenzyme complexes in the Booker, Boal, Cotruvo and Weinert labs. The ability to compliment the solution state AUC experiment with in-house multiangle light scattering, small angle X-ray scattering and isothermal calorimetry prior to high resolution X-ray crystallography and cryoEM studies would be a bonus. With this handshake, we are investing in true synergy where experimental results from the AUC would guide the structural interpretation and binding thermodynamics. The equipment will be used by eighteen structural biology faculty from two of our campuses, University Park and Hershey, and three colleges, College of Medicine, Eberly College of Science, and the College of Engineering. With a state-of-the-art multiwavelength Beckman Optima system, our goal is to ably resolve ultracentrifugation data in the spectral and hydrodynamic domains in a wide size range of applications, for a large number of users and enhance the capability at the Penn State X-ray crystallography core facility.