title: Comparison of fluorescence and light scattering based methods to assess formation and stability of protein-protein complexes.
abstract: Thermal shift methods such as differential scanning fluorimetry and differential static light scattering are widely used to identify stabilizing conditions for proteins that might promote crystallization. Here we report a comparison of the two methods when applied to optimization of buffer conditions for protein-protein complexes. Most of the protein complexes under study were amenable to analysis using these two techniques. Protein complexes behave towards thermal denaturation in a manner similar to single proteins, showing a more or less sharp transition consistent with a two-state model of unfolding. A comparison of the melting and aggregation temperatures for single components and the reconstituted complexes can provide additional evidence for complex formation and can be used to identify buffer conditions in which protein-protein complex formation is favored.
authors.affiliation: Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Scheele's Väg 2, S-17177 Stockholm, Sweden.
authors.forename: Jolanta
authors.initials: J
authors.lastname: Kopec
authors.forename: Gunter
authors.initials: G
authors.lastname: Schneider
mesh_terms: D002021:Buffers; D005470:Fluorometry / Q000379:methods*; D008027:Light; D046912:Multiprotein Complexes / Q000737:chemistry*; D011487:Protein Conformation; D011489:Protein Denaturation; D055503:Protein Multimerization; D055550:Protein Stability; D015203:Reproducibility of Results; D012542:Scattering, Radiation; D044366:Transition Temperature
chemical_list: D002021:Buffers; D046912:Multiprotein Complexes