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Portrait of Sara Snogerup Linse

Sara Linse

Professor

Portrait of Sara Snogerup Linse

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

Author

  • Wei-Feng Xue
  • J Carey
  • Sara Linse

Summary, in English

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 25degreesC, 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, DeltaGdegrees, of -42.5 kJ/mol. The enthalpic component, DeltaHdegrees, is -18.7 kJ/moI and the entropic contribution, DeltaSdegrees, is 79.8 J mol(-1) K-1 (-TDeltaSdegrees = -23.8 kJ/mol). The association of monellin is therefore a bimolecular intra-protein association whose energetics are slightly dominated by entropic factors. (C) 2004 Wiley-Liss, Inc.

Department/s

  • Biophysical Chemistry

Publishing year

2004

Language

English

Pages

586-595

Publication/Series

Proteins

Volume

57

Issue

3

Document type

Journal article

Publisher

John Wiley & Sons Inc.

Topic

  • Physical Chemistry

Status

Published

ISBN/ISSN/Other

  • ISSN: 0887-3585