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

Xue, Wei-Feng LU ; Carey, J and Linse, Sara LU (2004) In Proteins 57(3). p.586-595
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 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... (More)
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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Proteins
volume
57
issue
3
pages
586 - 595
publisher
John Wiley & Sons
external identifiers
  • pmid:15382228
  • wos:000224578400015
  • scopus:6344219892
ISSN
0887-3585
DOI
10.1002/prot.20241
language
English
LU publication?
yes
id
e105dd9f-f4e5-481d-a0eb-e4abf9c99c52 (old id 141063)
date added to LUP
2007-06-29 15:20:19
date last changed
2017-01-01 07:07:22
@article{e105dd9f-f4e5-481d-a0eb-e4abf9c99c52,
  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 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.},
  author       = {Xue, Wei-Feng and Carey, J and Linse, Sara},
  issn         = {0887-3585},
  language     = {eng},
  number       = {3},
  pages        = {586--595},
  publisher    = {John Wiley & Sons},
  series       = {Proteins},
  title        = {Multi-method global analysis of thermodynamics and kinetics in reconstitution of monellin},
  url          = {http://dx.doi.org/10.1002/prot.20241},
  volume       = {57},
  year         = {2004},
}