Improved methods for fitting sedimentation coefficient distributions derived by time-derivative techniques.
Time-derivative approaches to analyzing sedimentation velocity data have proven to be highly successful and have now been used routinely for more than a decade. For samples containing a small number of noninteracting species, the sedimentation coefficient distribution function, g(s *), traditionally has been fitted by Gaussian functions to derive the concentration, sedimentation coefficient, and diffusion coefficient of each species. However, the accuracy obtained by that approach is limited, even for noise-free data, and becomes even more compromised as more scans are included in the analysis to improve the signal/noise ratio (because the time span of the data becomes too large). Two new methods are described to correct for the effects of long time spans: one approach that uses a Taylor series expansion to correct the theoretical function and a second approach that creates theoretical g(s *) curves from Lamm equation models of the boundaries. With this second approach, the accuracy of the fitted parameters is approximately 0.1% and becomes essentially independent of the time span; therefore, it is possible to obtain much higher signal/noise when needed. This second approach is also compared with other current methods of analyzing sedimentation velocity data.
Philo
John S
JS
Alliance Protein Laboratories, Thousand Oaks, CA 91360, USA. jphilo@mailway.com
D000465:Algorithms; D003627:Data Interpretation, Statistical; D016018:Least-Squares Analysis; D015233:Models, Statistical; D011506:Proteins / Q000302:isolation & purification; D012984:Software; D013997:Time Factors; D014461:Ultracentrifugation / Q000706:statistics & numerical data*
D011506:Proteins