Sedimentation equilibrium analysis of mixed associations using numerical constraints to impose mass or signal conservation.

Publisher Summary This chapter describes the sedimentation equilibrium analysis of mixed associations, using numerical constraints to impose mass or signal conservation. Sedimentation equilibrium can be a very useful and powerful tool for characterizing the stoichiometry and strength of many physiologically important binding interactions. Many such interactions involve the binding of two or more different molecules (protein-protein, protein-ligand, protein-DNA, etc) and are, therefore, commonly termed “mixed” or “heterogeneous” associations. The chapter describes how to alleviate some of those difficulties through the use of numerical constraints during data analysis. Despite the intense interest in fields, such as signal transduction, which are governed by networks of specific binding interactions, the application of sedimentation equilibrium to characterize mixed associations is somewhat limited by difficulties in analyzing the data. An alternative to finding a means to determine the concentrations of individual components is to focus on these problems, with analyzing the total concentration data, by imposing mass conservation on the system.

[1]  T. Arakawa,et al.  Dimerization of the extracellular domain of granuloycte-colony stimulating factor receptor by ligand binding: a monovalent ligand induces 2:2 complexes. , 1996, Biochemistry.

[2]  T. Arakawa,et al.  Dimerization of the extracellular domain of the erythropoietin (EPO) receptor by EPO: one high-affinity and one low-affinity interaction. , 1996, Biochemistry.

[3]  J. Philo,et al.  Human Stem Cell Factor Dimer Forms a Complex with Two Molecules of the Extracellular Domain of Its Receptor, Kit (*) , 1996, The Journal of Biological Chemistry.

[4]  A. Minton,et al.  A strategy for efficient characterization of macromolecular heteroassociations via measurement of sedimentation equilibrium , 1991, Journal of molecular recognition : JMR.

[5]  M. Bubb,et al.  The interaction of monomeric actin with two binding sites on Acanthamoeba actobindin. , 1991, The Journal of biological chemistry.

[6]  T. Arakawa,et al.  Size-exclusion chromatography with on-line light-scattering, absorbance, and refractive index detectors for studying proteins and their interactions. , 1996, Analytical biochemistry.

[7]  M L Johnson,et al.  Analysis of data from the analytical ultracentrifuge by nonlinear least-squares techniques. , 1981, Biophysical journal.

[8]  T. Laue,et al.  Modern Analytical Ultracentrifugation , 1994, Emerging Biochemical and Biophysical Techniques.

[9]  A. Minton.  Alternative strategies for the characterization of associations in multicomponent solutions via measurement of sedimentation equilibrium , 1997 .

[10]  D. Winzor,et al.  Direct analysis of solute self-association by sedimentation equilibrium , 1996 .

[11]  T. Arakawa,et al.  Interactions of neurotrophin-3 (NT-3), brain-derived neurotrophic factor (BDNF), and the NT-3.BDNF heterodimer with the extracellular domains of the TrkB and TrkC receptors. , 1994, The Journal of biological chemistry.

[12]  G. Rivas,et al.  Rapid and accurate microfractionation of the contents of small centrifuge tubes: application in the measurement of molecular weight of proteins via sedimentation equilibrium. , 1993, Analytical biochemistry.

[13]  D. F. Senear,et al.  5-hydroxytryptophan as a new intrinsic probe for investigating protein-DNA interactions by analytical ultracentrifugation. Study of the effect of DNA on self-assembly of the bacteriophage lambda cI repressor. , 1993, Biochemistry.

[14]  M. S. Lewis,et al.  Ricin subunit association. Thermodynamics and the role of the disulfide bond in toxicity. , 1986, The Journal of biological chemistry.

[15]  G. Rivas,et al.  Sedimentation equilibrium-quantitative polyacrylamide gel electrophoresis (SE-QPAGE): a new technique for the detection of associations in multicomponent solutions. , 1993, Analytical biochemistry.

[16]  Michael L. Johnson,et al.  [16] Nonlinear least-squares analysis , 1985 .

[17]  J. Shiloach,et al.  Protein-protein interactions of HIV-1 reverse transcriptase: implication of central and C-terminal regions in subunit binding. , 1991, Biochemistry.

[18]  D. Millar,et al.  Dissociation equilibria of the tryptophan synthase α2β2 complex in saline buffer and guanidine isothiocyanate, as studied by sedimentation equilibrium , 1997 .