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

Xue, Wei-Feng, Carey, Jannette, Linse, Sara (2004) Multi-method global analysis of thermodynamics and kinetics in reconstitution of monellin. Proteins, 57 (3). pp. 586-95. ISSN 1097-0134. (doi:10.1002/prot.20241) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided)

The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided. (Contact us about this Publication)
Official URL
http://dx.doi.org/10.1002/prot.20241

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-protein association whose energetics are slightly dominated by entropic factors.

Item Type: Article
DOI/Identification number: 10.1002/prot.20241
Subjects: Q Science
Q Science > QC Physics
Q Science > QD Chemistry
Q Science > QP Physiology (Living systems) > QP517 Biochemistry
Divisions: Faculties > Sciences > School of Biosciences
Depositing User: Wei-Feng Xue
Date Deposited: 11 Jun 2013 12:49 UTC
Last Modified: 01 Aug 2019 10:35 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/34241 (The current URI for this page, for reference purposes)
Xue, Wei-Feng: https://orcid.org/0000-0002-6504-0404
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