1. We have simultaneously collected composition-dependent static and dynamic light scattering data on mixtures of non-interacting and interacting proteins, and developed a global analysis of the composition-dependence of both quantities. Contrary to expectation, addition of dynamic light scattering data did not enhance the resolution of characterization obtainable using static light scattering data only. Results of this work are being written up for publication. (B. Monterroso) 2. We have characterized the self-association equilibria of zinc-free insulin over a wide range of pH values, via measurement of the concentration dependence of static light scattering (A. Attri). At pH 1.6, insulin is a non-associating monomer in acetate buffer and a very weakly self-associating dimer in HCl. At pH values between 3 and 8, the concentration dependence of scattering is quantitatively accounted for by a simple isodesmic indefinite association scheme. At pH 10, the concentration dependence of scattering is quantitatively accounted for by a modified isodesmic scheme in which the equilibrium association constant for addition of monomer to monomer is about five times smaller than the equilibrium association constant for addition of monomer to all higher oligomers. A report of this work has been published (A. Attri, C. Fernandez). 3. We have characterized the self-association equilibria of insulin in the presence of zinc. Analysis of the concentration dependence of static light scattering indicates that over the concentration range studied (0.3 - 5 mg/ml), insulin behaves as a hexamer undergoing isodesmic indefinite self-association. Analysis of the concentration dependence of the dynamic light scattering confirms this interpretation. A report of this work has been published (A. Attri, C. Fernandez). 4. We have characterized the interaction between tau protein and a 7K molecular weight fraction of heparin, a sulfated polysaccharide with high negative charge density. The composition dependence of the scattering intensity is well described by a simple 1:1 heteroassociation. This is the first application of composition gradient - static light scattering to the association between two different types of macromolecule. A report of this work has been published (C. Fernandez, Y. Liang). 5. We have measured the dependence of the intensity of light scattering upon protein concentration in solutions of FtsZ containing either GTP an a GTP-regenerating system or GPCPP, a slowly hydrolyzable GTP analog, in the presence of varying concentrations of Mg++ ion. The simplest interpretation of the observed dependence is that self-association of either FtsZ-GTP or FtsZ-GPCPP proceeds initially by linear growtn according to an indefinite isodesmic scheme, followed by cooperative formation of a stable oligomer or narrow size distribution of oligomers of size 30-40 subunits that are thought to be cyclic. This work is being written up for publication (G. Rivas).

Project Start
Project End
Budget Start
Budget End
Support Year
4
Fiscal Year
2010
Total Cost
$306,700
Indirect Cost
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State
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Wu, Di; Minton, Allen P (2015) Quantitative characterization of nonspecific self- and hetero-interactions of proteins in nonideal solutions via static light scattering. J Phys Chem B 119:1891-8
Wu, Di; Minton, Allen P (2013) Compensating effects of urea and trimethylamine-N-oxide on the heteroassociation of ?-chymotrypsin and soybean trypsin inhibitor. J Phys Chem B 117:3554-9
Wu, Di; Minton, Allen P (2013) Quantitative characterization of the compensating effects of trimethylamine-N-oxide and guanidine hydrochloride on the dissociation of human cyanmethmoglobin. J Phys Chem B 117:9395-9
Wu, Di; Minton, Allen P (2013) Quantitative characterization of the interaction between sucrose and native proteins via static light scattering. J Phys Chem B 117:111-7
Monterroso, Begoña; Ahijado-Guzmán, Rubén; Reija, Belén et al. (2012) Mg(2+)-linked self-assembly of FtsZ in the presence of GTP or a GTP analogue involves the concerted formation of a narrow size distribution of oligomeric species. Biochemistry 51:4541-50
Monterroso, Begoña; Rivas, Germán; Minton, Allen P (2012) An equilibrium model for the Mg(2+)-linked self-assembly of FtsZ in the presence of GTP or a GTP analogue. Biochemistry 51:6108-13
Fernández, Cristina; Minton, Allen P (2011) Effect of nonadditive repulsive intermolecular interactions on the light scattering of concentrated protein-osmolyte mixtures. J Phys Chem B 115:1289-93
Rivas, Germán; Minton, Allen P (2011) Beyond the second virial coefficient: Sedimentation equilibrium in highly non-ideal solutions. Methods 54:167-74
Zhu, Hai-Li; Fernandez, Cristina; Fan, Jun-Bao et al. (2010) Quantitative characterization of heparin binding to Tau protein: implication for inducer-mediated Tau filament formation. J Biol Chem 285:3592-9
Scherer, Thomas M; Liu, Jun; Shire, Steven J et al. (2010) Intermolecular interactions of IgG1 monoclonal antibodies at high concentrations characterized by light scattering. J Phys Chem B 114:12948-57

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