Multi-speed sedimentation velocity simulations with UltraScan-III

European Biophysics Journal : EBJ
Tayler L WilliamsBorries Demeler

Abstract

Recent developments in the UltraScan-III software make it possible to model multi-speed analytical ultracentrifugation sedimentation velocity experiments using finite-element solutions of the Lamm equation. Using simulated data, we demonstrate here how these innovations can be used to enhance the resolution of sedimentation velocity experiments when compared to single-speed experiments. Using heterogeneous systems covering as much as five orders of magnitude in molar mass and fivefold in anisotropy, we compare results from runs performed at multiple speeds to those obtained from single-speed experiments, fitted individually and analyzed globally over multiple speeds, and quantify resolution for sample heterogeneous in size and anisotropy. We also provide guidance on the design of multi-speed experiments and offer a program that can be used to deduce optimal spacing of rotor speeds and speed step durations when a few parameters from the experiment can be estimated. These include the meniscus position, the sedimentation coefficient of the largest species in a mixture, and a solute distribution. Our results show that errors observed in the determination of hydrodynamic parameters for system with great heterogeneity are markedly re...Continue Reading

References

Sep 29, 2015·Methods in Enzymology·Ryan A Rogge, Jeffrey C Hansen
Apr 4, 2018·European Biophysics Journal : EBJ·Gary E GorbetBorries Demeler

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Citations

Sep 16, 2018·European Biophysics Journal : EBJ·Olwyn ByronTrushar R Patel
Nov 26, 2020·European Biophysics Journal : EBJ·Marielle StoutjesdykBorries Demeler

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Software Mentioned

UltraScan
MC
Proteomelab
Optima
III
2DSA
GA
Optima AUC

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