title: Energetics of subunit dimerization in bacteriophage lambda cI repressor: linkage to protons, temperature, and KCl.
abstract: A common feature of gene regulatory systems is the linkage between reversible protein oligomerization and DNA binding. Experimental dissection using temperature dependence of the subunit-subunit energetics and their linkage to processes such as ion binding and release is necessary for characterization of the chemical forces that contribute to cooperativity and site specificity. We have therefore studied the effects of temperature, proton activity, and monovalent salt on monomer-dimer assembly of the lambda cI repressor using a recently developed gel chromatographic procedure. This technique has made possible studies in the previously inaccessible picomolar concentration ranges where the assembly reactions occur. Upon formation of the dimer interface in the range pH 5-9, we find an overall absorption of protons which is temperature-dependent. The dimerization reaction displays a large negative enthalpy of association at all conditions studied (pH 5, 7, and 9). The reaction is also dependent on monovalent salt concentration: subunit association is weaker at low-salt conditions. The results suggest that a repulsive interaction between negatively charged side chains (i.e., aspartates and glutamates) on each monomer surface is attenuated by increasing concentrations of KCl. Formation of the dimer interface may be mediated by absorption of cations which stabilize the complex.
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, Missouri 63110.
authors.forename: K S
authors.initials: KS
authors.lastname: Koblan
authors.forename: G K
authors.initials: GK
authors.lastname: Ackers
mesh_terms: D010582:Bacteriophage lambda / Q000378:metabolism*; D002151:Calorimetry; D004268:DNA-Binding Proteins; D004926:Escherichia coli / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D046911:Macromolecular Substances; D008433:Mathematics; D009994:Osmolar Concentration; D011189:Potassium Chloride / Q000494:pharmacology*; D012097:Repressor Proteins / Q000737:chemistry / Q000378:metabolism*; D013816:Thermodynamics; D014157:Transcription Factors / Q000378:metabolism; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins
chemical_list: D004268:DNA-Binding Proteins; D046911:Macromolecular Substances; D012097:Repressor Proteins; D014157:Transcription Factors; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; C016649:phage repressor proteins; D011189:Potassium Chloride