An improved approximate solution of the Lamm equation for the simultaneous estimation of sedimentation and diffusion coefficients from sedimentation velocity experiments

• Published in: Biophysical chemistry 1998-02, Vol.70 (2), p.133-146
• Main Authors: Behlke, Joachim, Ristau, Otto
• Format: Article
• Language: English
• Subjects: Angiotensin; Diffusion; Gross conformation; Lamm equation; Nonlinear fit; Sedimentation
• ISSN: 0301-4622; 1873-4200
• DOI: 10.1016/S0301-4622(97)00124-5

Sedimentation and diffusion coefficients are important parameters to describe size and shape of macromolecules in solution. The data can be obtained from sedimentation velocity experiments by a nonlinear fitting procedure using approximate solutions for the Lamm equation. Here, we present a modification of such a model function that was originally proposed by Fujita [H. Fujita, Mathematical Theory of Sedimentation Analysis, Wiley, New York, 1962]. The extended model function is well suitable to study low molecular mass compounds. The improvement of this solution given here is based on using an adjustable value for the explicit integration variable, z, the reduced radius. This modification leads to more accurate sedimentation and diffusion coefficients compared to using a constant value of 0.5 as used by Fujita. The advantage of our modification was demonstrated by the analysis of noise-free curves calculated using the finite element method, as well as experimental curves obtained for the peptides angiotensin I and II. The relatively low sedimentation and diffusion coefficients found for both substances indicate that the peptides exist as extended chains of about 3.65 nm (angiotensin I) or 3.04 nm length (angiotensin II) in solution. The lack of higher-order structure of the peptides that was derived also from CD spectra might facilitate receptor binding, and could be one reason for the fast proteolytic digestion of the free peptides.