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Influence of complexing polyanions on the thermostability of basic protiens.

Ivinova, Olga LU ; Izumrudov, Vladimir LU ; Muronetz, V; Galaev, Igor LU and Mattiasson, Bo LU (2003) In Macromolecular Bioscience 3(3-4). p.210-215
Abstract
Lysozyme (Lyz), chymotrypsinogen (Cht), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were used as model proteins capable of forming water-soluble polyelectrolyte complexes with linear synthetic polyanions. The complex formation with sodium poly(methacrylate) (PMA), sodium poly(acrylate) (PAA), sodium poly(anetholsulfonate) (PAS), and potassium poly(vinylsulfate) (PVS) markedly reduced the temperature of protein denaturation, Tmax, as determined by differential scanning calorimetry (DSC). The effect of sodium poly(styrenesulfonate) (PSS) on Lyz was so drastic that the protein melting peak was not observed at all during DSC measurements. The temperature shift, most pronounced for Lyz, increased upon substitution of the polyanions... (More)
Lysozyme (Lyz), chymotrypsinogen (Cht), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were used as model proteins capable of forming water-soluble polyelectrolyte complexes with linear synthetic polyanions. The complex formation with sodium poly(methacrylate) (PMA), sodium poly(acrylate) (PAA), sodium poly(anetholsulfonate) (PAS), and potassium poly(vinylsulfate) (PVS) markedly reduced the temperature of protein denaturation, Tmax, as determined by differential scanning calorimetry (DSC). The effect of sodium poly(styrenesulfonate) (PSS) on Lyz was so drastic that the protein melting peak was not observed at all during DSC measurements. The temperature shift, most pronounced for Lyz, increased upon substitution of the polyanions according to the following series: PMA < PVS < PAA < PAS < PSS. Decomposition of the complexes by addition of either sodium chloride or poly(N-ethyl-4-vinylpyridinium) cation completely restored the initial Tmax of the protein (except for PSS and PAS). The complex formation slightly affected the enzyme activity up to temperatures close to Tmax of the polyanion-protein complex. On further heating, the activity of the complex decreased steeply, whereas the free enzyme maintained a high activity. The data obtained strongly suggest that the protein-polyelectrolyte interactions in solution, while leaving the thermostability and activity of the proteins practically unaffected over a rather wide temperature range, result in the effective denaturation of proteins once a certain critical temperature is achieved. This finding appears to be crucial for further development of immobilized enzymes in biotechnology and essential for understanding mechanisms and principles of the functioning of proteins immobilized on charged matrices in vivo. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Macromolecular Bioscience
volume
3
issue
3-4
pages
210 - 215
publisher
John Wiley & Sons
external identifiers
  • wos:000182698000008
ISSN
1616-5195
DOI
language
English
LU publication?
yes
id
f3da843b-cf89-4349-9b5d-ec1f23512cf9 (old id 129145)
date added to LUP
2007-07-02 11:45:54
date last changed
2018-05-29 11:42:35
@article{f3da843b-cf89-4349-9b5d-ec1f23512cf9,
  abstract     = {Lysozyme (Lyz), chymotrypsinogen (Cht), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were used as model proteins capable of forming water-soluble polyelectrolyte complexes with linear synthetic polyanions. The complex formation with sodium poly(methacrylate) (PMA), sodium poly(acrylate) (PAA), sodium poly(anetholsulfonate) (PAS), and potassium poly(vinylsulfate) (PVS) markedly reduced the temperature of protein denaturation, Tmax, as determined by differential scanning calorimetry (DSC). The effect of sodium poly(styrenesulfonate) (PSS) on Lyz was so drastic that the protein melting peak was not observed at all during DSC measurements. The temperature shift, most pronounced for Lyz, increased upon substitution of the polyanions according to the following series: PMA &lt; PVS &lt; PAA &lt; PAS &lt; PSS. Decomposition of the complexes by addition of either sodium chloride or poly(N-ethyl-4-vinylpyridinium) cation completely restored the initial Tmax of the protein (except for PSS and PAS). The complex formation slightly affected the enzyme activity up to temperatures close to Tmax of the polyanion-protein complex. On further heating, the activity of the complex decreased steeply, whereas the free enzyme maintained a high activity. The data obtained strongly suggest that the protein-polyelectrolyte interactions in solution, while leaving the thermostability and activity of the proteins practically unaffected over a rather wide temperature range, result in the effective denaturation of proteins once a certain critical temperature is achieved. This finding appears to be crucial for further development of immobilized enzymes in biotechnology and essential for understanding mechanisms and principles of the functioning of proteins immobilized on charged matrices in vivo.},
  author       = {Ivinova, Olga and Izumrudov, Vladimir and Muronetz, V and Galaev, Igor and Mattiasson, Bo},
  issn         = {1616-5195},
  language     = {eng},
  number       = {3-4},
  pages        = {210--215},
  publisher    = {John Wiley & Sons},
  series       = {Macromolecular Bioscience},
  title        = {Influence of complexing polyanions on the thermostability of basic protiens.},
  url          = {http://dx.doi.org/},
  volume       = {3},
  year         = {2003},
}