Molecular mass determination by sedimentation velocity experiments and direct fitting of the concentration profiles
| Autor: | J. Behlke, O. Ristau |
|---|---|
| Přispěvatelé: | MDC Library |
| Rok vydání: | 1997 |
| Předmět: |
Cancer Research
Analytical chemistry Biophysics Thermodynamics 570 Life Sciences 610 Medical Sciences Medicine Diffusion Animals Computer Simulation Centrifugation Diffusion (business) Theoretical Models Molecular mass Myoglobin Chemistry Whales Proteins Lamm equation Function (mathematics) Models Theoretical Sedimentation Molecular Weight Sedimentation coefficient Kinetics Spectrophotometry Non-linear least squares Muramidase Chickens Ultracentrifugation Software Research Article |
| Zdroj: | Biophysical Journal 72 (1): 428-434. |
| ISSN: | 0006-3495 |
| DOI: | 10.1016/s0006-3495(97)78683-1 |
| Popis: | A new method for the direct molecular mass determination from sedimentation velocity experiments is presented. It is based on a nonlinear least squares fitting procedure of the concentration profiles and simultaneous estimation of the sedimentation and diffusion coefficients using approximate solutions of the Lamm equation. A computer program, LAMM, was written by using five different model functions derived by Fujita (1962, 1975) to describe the sedimentation of macromolecules during centrifugation. To compare the usefulness of these equations for the analysis of hydrodynamic results, the approach was tested on data sets of Claverie simulations as well as experimental curves of some proteins. A modification for one of the model functions is suggested, leading to more reliable sedimentation and diffusion coefficients estimated by the fitting procedure. The method seems useful for the rapid molecular mass determination of proteins larger than 10 kDa. One of the equations of the Archibald type is also suitable for compounds of low molecular mass, probably less than 10 kDa, because this model function requires neither the plateau region nor a meniscus free of solute. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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