Rapid determination of molar mass in modified Archibald experiments using direct fitting of the Lamm equation.
A new method is described that allows measurement of the molar mass of the solute within 15 to 30 min after start of a conventional long-column sedimentation equilibrium experiment. A series of scans of the concentration distribution in close vicinity of the meniscus, taken in rapid succession after the start of the centrifuge run, is analyzed by direct fitting using the Lamm equation and the Svedberg equation. In case of a single solute, this analysis of the initial depletion at the meniscus reveals its buoyant molar mass and sedimentation coefficient with an accuracy of approximately 10% and provides gross information about sample heterogeneity. This method can be used to study macromolecules that do not possess the prolonged stability needed in conventional sedimentation equilibrium experiments and it can increase the efficiency of sedimentation equilibrium experiments of previously uncharacterized samples.
Schuck
P
P
Molecular Interactions Resource, BEPS, OD, National Institutes of Health, Bethesda, Maryland 20892, USA. pschuck@helix.nih.gov
Millar
D B
DB
D002621:Chemistry / Q000379:methods*; D004798:Enzymes / Q000737:chemistry; D008962:Models, Theoretical; D008970:Molecular Weight; D011506:Proteins / Q000737:chemistry*; D012996:Solutions; D013816:Thermodynamics; D013997:Time Factors
D004798:Enzymes; D011506:Proteins; D012996:Solutions