A good starting place: Biophys J. 1996 Oct;71(4):2049-55. Evaluation of linked protonation effects in protein binding reactions using isothermal titration calorimetry. Baker BM, Murphy KP.
Abstract A theoretical development in the evaluation of proton linkage in protein binding reactions by isothermal titration calorimetry (ITC) is presented. For a system in which binding is linked to protonation of an ionizable group on a protein, we show that by performing experiments as a function of pH in buffers with varying ionization enthalpy, one can determine the pK(a)'s of the group responsible for the proton linkage in the free and the liganded states, the protonation enthalpy for this group in these states, as well as the intrinsic energetics for ligand binding (delta H(o), delta S(o), and delta C(p)). Determination of intrinsic energetics in this fashion allows for comparison with energetics calculated empirically from structural information. It is shown that in addition to variation of the ligand binding constant with pH, the observed binding enthalpy and heat capacity change can undergo extreme deviations from their intrinsic values, depending upon pH and buffer conditions. PMID: 8889179 [PubMed - indexed for MEDLINE] PMCID: PMC1233671 On Apr 4, 2013, at 8:56 AM, Deepak Oswal wrote: Dear colleagues: I was wondering if you could kindly share your thoughts and help me understand the relationship between pKa and affinity of a protein for a ligand. Are these two properties related? Specifically, does a lysine with a pKa of 8.5 have a greater affinity for a negatively charged ligand than a lysine with a pKa of 10.5 for the same ligand at physiological pH? Any comments would be highly appreciated. Deepak
