1. The concentration dependence of light scattering of three proteins, ovalbumin, ribonuclease A, and ovomucoid, and binary mixtures of the three proteins were measured at various compositions and pH values at total protein concentrations of up to 35 g/l, under conditions such that ribonuclease bears a net positive charge and the other proteins bear a net negative charge. The concentration dependence was interpreted in the context of three models: a statistical thermodynamic model and two extensions of the effective hard particle model generalized to allow for the presence of non-additive interactions between species in a binary mixture (D. Wu, A. Minton). 2. Experiments were performed to determine whether quantitative composition gradient light scattering experiments can be performed using a fluorescence spectrophotometer in place of a dedicated light scattering photometer. Results obtained so far are encouraging, but further work will be required in order to achieve the requisite reproducibility.

Project Start
Project End
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Budget End
Support Year
8
Fiscal Year
2014
Total Cost
Indirect Cost
Name
U.S. National Inst Diabetes/Digst/Kidney
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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
Attri, Arun K; Fernández, Cristina; Minton, Allen P (2010) pH-dependent self-association of zinc-free insulin characterized by concentration-gradient static light scattering. Biophys Chem 148:28-33
Attri, Arun K; Fernández, Cristina; Minton, Allen P (2010) Self-association of Zn-insulin at neutral pH: investigation by concentration gradient--static and dynamic light scattering. Biophys Chem 148:23-7

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