Multi-method global analysis of thermodynamics and kinetics in reconstitution of monellin

Proteins
Wei-Feng XueSara Linse

Abstract

Accurate and precise determinations of thermodynamic parameters of binding are important steps toward understanding many biological mechanisms. Here, a multi-method approach to binding analysis is applied and a detailed error analysis is introduced. Using this approach, the binding thermodynamics and kinetics of the reconstitution of the protein monellin have been quantitatively determined in detail by simultaneous analysis of data collected with fluorescence spectroscopy, surface plasmon resonance and isothermal titration calorimetry at 25 degrees C, pH 7.0 and 150 mM NaCl. Monellin is an intensely sweet protein composed of two peptide chains that form a single globular domain. The kinetics of the reconstitution reaction are slow, with an association rate constant, k(on) of 8.8 x 10(3) M(-1) s(-1) and a dissociation rate constant, k(off) of 3.1 x 10(-4) s(-1). The equilibrium constant K(A) is 2.8 x 10(7) M(-1) corresponding to a standard free energy of association, DeltaG degrees , of -42.5 kJ/mol. The enthalpic component, DeltaH degrees , is -18.7 kJ/mol and the entropic contribution, DeltaS degrees , is 79.8 J mol(-1) K(-1) (-TDeltaS degrees = -23.8 kJ/mol). The association of monellin is therefore a bimolecular intra-protei...Continue Reading

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Citations

May 27, 2009·International Journal of Molecular Sciences·Mikael C BauerSara Linse
Dec 23, 2008·Biochimica Et Biophysica Acta·Wei-Feng XueJannette Carey
Oct 27, 2007·Protein Science : a Publication of the Protein Society·Jannette CareySara Linse
Oct 13, 2005·Journal of Molecular Recognition : JMR·Abdessamad Ababou, John E Ladbury
Nov 10, 2010·Protein Expression and Purification·Nilesh Aghera, Jayant B Udgaonkar
Mar 1, 2008·Journal of Molecular Biology·Anders SandbergTorleif Härd
Apr 28, 2009·Journal of Molecular Biology·Ashish K Patra, Jayant B Udgaonkar
Jan 31, 2006·Biophysical Journal·Stina LindmanSara Linse
Oct 28, 2005·Journal of Molecular Recognition : JMR·Rebecca L Rich, David G Myszka
Aug 26, 2018·Scientific Reports·Tanja Weiffert, Sara Linse
Nov 16, 2010·Molecular BioSystems·Olga SzczepankiewiczSara Linse

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