title: Counteracting osmolyte trimethylamine N-oxide destabilizes proteins at pH below its pKa. Measurements of thermodynamic parameters of proteins in the presence and absence of trimethylamine N-oxide.
abstract: Earlier studies have reported that trimethylamine N-oxide (TMAO), a naturally occurring osmolyte, is a universal stabilizer of proteins because it folds unstructured proteins and counteracts the deleterious effects of urea and salts on the structure and function of proteins. This conclusion has been reached from the studies of the effect of TMAO on proteins in the pH range 6.0-8.0. In this pH range TMAO is almost neutral (zwitterionic form), for it has a pK(a) of 4.66 +/- 0.10. We have asked the question of whether the effect of TMAO on protein stability is pH-dependent. To answer this question we have carried out thermal denaturation studies of lysozyme, ribonuclease-A, and apo-alpha-lactalbumin in the presence of various TMAO concentrations at different pH values above and below the pK(a) of TMAO. The main conclusion of this study is that near room temperature TMAO destabilizes proteins at pH values below its pK(a), whereas it stabilizes proteins at pH values above its pK(a). This conclusion was reached by determining the T(m) (midpoint of denaturation), delta H(m) (denaturational enthalpy change at T(m)), delta C(p) (constant pressure heat capacity change), and delta G(D) degrees (denaturational Gibbs energy change at 25 degrees C) of proteins in the presence of different TMAO concentrations. Other conclusions of this study are that T(m) and delta G(D) degrees depend on TMAO concentration at each pH value and that delta H(m) and the delta C(p) are not significantly changed in presence of TMAO.
authors.affiliation: Department of Biosciences, Jamia Millia Islamia, Jamia Nagar, New Delhi 110 025, India.
authors.forename: Rajendrakumar
authors.initials: R
authors.lastname: Singh
authors.forename: Inamul
authors.initials: I
authors.lastname: Haque
authors.forename: Faizan
authors.initials: F
authors.lastname: Ahmad
mesh_terms: D006863:Hydrogen-Ion Concentration; D008744:Methylamines / Q000494:pharmacology*; D011506:Proteins / Q000737:chemistry*; D013816:Thermodynamics
chemical_list: D008744:Methylamines; D011506:Proteins; C005855:trimethyloxamine