New methods for measuring macromolecular interactions in solution via static light scattering: basic methodology and application to nonassociating and self-associating proteins.
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).
Attri
Arun K
AK
Section on Physical Biochemistry, Laboratory of Biochemistry and Genetics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, U.S. Department of Health and Human Services, Bethesda, MD 20892, USA.
Minton
Allen P
AP
D019281:Dimerization; D004867:Equipment Design; D008027:Light; D008962:Models, Theoretical; D046912:Multiprotein Complexes / Q000032:analysis*; D011506:Proteins / Q000737:chemistry*; D012542:Scattering, Radiation; D012996:Solutions
D046912:Multiprotein Complexes; D011506:Proteins; D012996:Solutions