A calorimetric investigation of the interaction of the lac repressor with inducer.
A calorimetric study has been made of the interaction between the lac repressor and isopropyl-1-thio-beta-D-galactopyranoside (IPTG). The buffer-corrected enthalpy of reaction at 25 degrees C was found to be -15.6, -24.7, -4.6 kJ/mol of bound IPTG at pH 7.0, pH 8.1, and pH 9.0, respectively. This large range of enthalpy values is in contrast to a maximum difference in the free energy of the reaction of only 1.5 kJ/mol of bound IPTG between these pH values. The reaction was found by calorimetric measurements in different buffers to be accompanied by an uptake of 0.29 mol of protons/mol of bound IPTG at pH 8.1. The pH dependency of the reaction enthalpy suggests differences in the extent of protonation of the binding site and the involvement of H bonding with IPTG. The lack of strong hydrophobic contributions in the IPTG binding process is revealed by the absence of any determinable heat capacity change for the reaction at pH 7.0. The presence of phosphate buffer significantly alters the enthalpy of IPTG binding at higher pH values, but has little effect upon the binding constant. This implies that highly negative phosphate species change the nature of the IPTG binding site without any displacement of phosphate upon IPTG binding.
D002021:Buffers; D002151:Calorimetry; D004926:Escherichia coli / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D007700:Kinetics; D010957:Plasmids; D012097:Repressor Proteins / Q000378:metabolism*; D013865:Thioglycosides / Q000494:pharmacology; D014157:Transcription Factors / Q000378:metabolism*
D002021:Buffers; D012097:Repressor Proteins; D013865:Thioglycosides; D014157:Transcription Factors; D007544:Isopropyl Thiogalactoside