New methods for measuring macromolecular interactions in solution via static light scattering: basic methodology and application to nonassociating and self-associating proteins.

Arun K Attri; Allen P Minton

doi:10.1016/j.ab.2004.09.045

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New methods for measuring macromolecular interactions in solution via static light scattering: basic methodology and application to nonassociating and self-associating proteins.

Arun K Attri·Allen P Minton

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Abstract

A method for rapid detection and characterization of reversible associations of macromolecules in solution is presented. A programmable dual-syringe infusion pump is used to introduce a solution of time-varying composition into parallel flow cells for concurrent measurement of laser light scattering at multiple angles and ultraviolet-visible absorbance. An experiment lasting less than 15 min produces a large and information-rich set of data, consisting of several thousand values of the Rayleigh ratio as a function of solute concentration(s) and scattering angle. Using a novel treatment of the data, the entire data set may be equally rapidly analyzed in the context of models for self-association. Validation experiments conducted on previously characterized nonassociating and self-associating proteins yielded robust values for molecular weights in the range 10-330 kDa and equilibrium association constants for dimer formation in the range 2 x 10(3)-6 x 10(5) M(-1).

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Dimerization Equipment Design Light Models, Theoretical Multiprotein Complexes Proteins Scattering, Radiation Solutions Journal Article Research Support, U.S. Gov't, P.H.S.

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Arun K Attri, Allen P Minton. 2005-02-01. New methods for measuring macromolecular interactions in solution via static light scattering: basic methodology and application to nonassociating and self-associating proteins.. https://doi.org/10.1016/j.ab.2004.09.045

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