Multi-method global analysis of thermodynamics and kinetics in reconstitution of monellin
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/141063
- author
- Xue, Wei-Feng LU ; Carey, J and Linse, Sara LU
- organization
- publishing date
- 2004
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Proteins
- volume
- 57
- issue
- 3
- pages
- 586 - 595
- publisher
- John Wiley & Sons Inc.
- 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
- 2016-04-01 16:30:42
- date last changed
- 2022-01-28 20:12:22
@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 Inc.}},
series = {{Proteins}},
title = {{Multi-method global analysis of thermodynamics and kinetics in reconstitution of monellin}},
url = {{http://dx.doi.org/10.1002/prot.20241}},
doi = {{10.1002/prot.20241}},
volume = {{57}},
year = {{2004}},
}