Simultaneous rapid estimation of sedimentation coefficient and molecular weight.
Data obtained from the early portion of sedimentation velocity experiments may be analyzed to simultaneously estimate both s and s/D. The C versus r data obtained are analyzed using a nonlinear least squares algorithm and an approximate solution to the Lamm equation. This procedure was tested both with simulated noisy data and with experimental data obtained using ribonuclease, ovalbumin, and somatostatin.dodecylsulfate. The procedure assumes that both s and D are independent of concentration. The results suggest that optimal estimation of both s and s/D is obtained at values of (= 2D/(s.omega(2).r(a)(2))) in the range of 0.002 to 0.01 and values of tau(= 2 somega(2)t) less than 0.04. Appropriate selection of rotor speed allows the estimation of both s and s/D for nearly globular macromolecules in the range of 10(4) to 10(6) daltons with data obtained during the first 3000-5000 seconds of a sedimentation velocity experiment.
Holladay
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Department of Biochemistry, Vanderbilt University, Nashville, Tennessee 37232, USA.