Abstract
An increasingly common question in structural biology now is the state of aggregation of a protein or complex in solution. In other words, is it monomeric, or has it aggregated to form some oligomer which is essential for activity? One approach to answering this question has been the use of gel permeation chromatography, where a porous gel matrix is packed into a column and the size of the complex measured in terms of its elution volume. The column is calibrated for elution volumes by using a set of standard proteins, of known molecular mass, and the mass of the unknown is estimated from a semi-logarithmic plot of molecular mass against elution volume. There is a problem with this approach, namely that the elution volume is actually a function of Stokes radius, rather than molecular mass, and so the calibration is based on the, often unacknowledged, assumption that the molecular shapes of the unknown and standards will be similar.
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Jonathan, P., Butler, G. (2001). Analytical Centrifugation: Looking at Aggregation in Free Solution. In: Pifat-Mrzljak, G. (eds) Supramolecular Structure and Function 7. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-1363-6_2
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DOI: https://doi.org/10.1007/978-1-4615-1363-6_2
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