Sedimentation velocity method in the analytical ultracentrifuge for the study of protein-protein interactions.
Sedimentation analysis in the analytical ultracentrifuge can be employed to detect macromolecular interactions. Whenever two molecules interact the mass of the resulting complex is increased and this is reflected in the sedimentation behavior. In this chapter we discuss how this phenomenon can be utilized to determine quantitative parameters of an interaction. An example, interaction of single-stranded DNA binding protein with a subunit of DNA polymerase III holoenzyme is given together with a thorough treatment of the relating theory and a description of evaluation algorithms.
Urbanke
Claus
C
Department of Biophysical Chemistry, Medizinische Hochschule, Hannover, Germany.
Witte
Gregor
G
Curth
Ute
U
D000465:Algorithms; D004258:DNA Polymerase III / Q000737:chemistry / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D008954:Models, Biological; D046912:Multiprotein Complexes / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D021122:Protein Subunits; D014461:Ultracentrifugation / Q000295:instrumentation / Q000379:methods* / Q000706:statistics & numerical data
D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D008024:Ligands; D046912:Multiprotein Complexes; D021122:Protein Subunits; C476430:SSB protein, E coli; D004258:DNA Polymerase III