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Protein stabilization by compatible solutes - Effect of diglycerol phosphate on the dynamics of Desulfovibrio gigas rubredoxin studied by NMR

Protein stabilization by compatible solutes - Effect of diglycerol phosphate on the dynamics of Desulfovibrio gigas rubredoxin studied by NMR
Protein stabilization by compatible solutes - Effect of diglycerol phosphate on the dynamics of Desulfovibrio gigas rubredoxin studied by NMR
Heteronuclear NMR relaxation measurements and hydrogen exchange data have been used to characterize protein dynamics in the presence or absence of stabilizing solutes from hyperthermophiles. Rubredoxin from Desulfovibrio gigas was selected as a model protein and the effect of diglycerol phosphate on its dynamic behaviour was studied. The presence of 100 mM diglycerol phosphate induces a fourfold increase in the half-life for thermal denaturation of D. gigas rubredoxin [Lamosa, P., Burke, A., Peist, R., Huber, R., Liu, M.Y., Silva, G., Rodrigues-Pousada, C., LeGall, J., Maycock, C. & Santos, H. (2000) Appl. Environ. Microbiol.66, 1974-1979]. A model-free analysis of the protein backbone relaxation parameters shows an average increase of generalized order parameters of 0.015 reflecting a small overall reduction in mobility of fast-scale motions. Hydrogen exchange data acquired over a temperature span of 20degreesC yielded thermodynamic parameters for the structural opening reactions that allow for the exchange. This shows that the closed form of the protein is stabilized by an additional 1.6 kJ.mol(-1) in the presence of the solute. The results seem to indicate that the stabilizing effect is due mainly to a reduction in mobility of the slower, larger-scale motions within the protein structure with an associated increase in the enthalpy of interactions.
chemical exchange, compatible solutes, protein dynamics, rubredoxin, thermostability, backbone dynamics, hydrogen-exchange, preferential interactions, temperature-dependence, globular-proteins, organic solutes, relaxation, stability, binding, spectroscopy
0014-2956
4606-4614
Lamosa, Pedro
9c2fdf0a-8bc3-4ed6-b295-f26bc448bfd7
Turner, David L.
6c4c1ccb-a6f2-47cf-944e-e69269198a29
Ventura, Rita
fa67726f-3b7d-4ae9-9100-20f29b7a58ff
Maycock, Christopher
b45ba495-16b5-4529-9b8e-322796828e37
Santos, Helena
1c7cb520-ed15-41e7-b381-467ca075ed30
Lamosa, Pedro
9c2fdf0a-8bc3-4ed6-b295-f26bc448bfd7
Turner, David L.
6c4c1ccb-a6f2-47cf-944e-e69269198a29
Ventura, Rita
fa67726f-3b7d-4ae9-9100-20f29b7a58ff
Maycock, Christopher
b45ba495-16b5-4529-9b8e-322796828e37
Santos, Helena
1c7cb520-ed15-41e7-b381-467ca075ed30

Lamosa, Pedro, Turner, David L., Ventura, Rita, Maycock, Christopher and Santos, Helena (2003) Protein stabilization by compatible solutes - Effect of diglycerol phosphate on the dynamics of Desulfovibrio gigas rubredoxin studied by NMR. European Journal of Biochemistry, 270 (23), 4606-4614.

Record type: Article

Abstract

Heteronuclear NMR relaxation measurements and hydrogen exchange data have been used to characterize protein dynamics in the presence or absence of stabilizing solutes from hyperthermophiles. Rubredoxin from Desulfovibrio gigas was selected as a model protein and the effect of diglycerol phosphate on its dynamic behaviour was studied. The presence of 100 mM diglycerol phosphate induces a fourfold increase in the half-life for thermal denaturation of D. gigas rubredoxin [Lamosa, P., Burke, A., Peist, R., Huber, R., Liu, M.Y., Silva, G., Rodrigues-Pousada, C., LeGall, J., Maycock, C. & Santos, H. (2000) Appl. Environ. Microbiol.66, 1974-1979]. A model-free analysis of the protein backbone relaxation parameters shows an average increase of generalized order parameters of 0.015 reflecting a small overall reduction in mobility of fast-scale motions. Hydrogen exchange data acquired over a temperature span of 20degreesC yielded thermodynamic parameters for the structural opening reactions that allow for the exchange. This shows that the closed form of the protein is stabilized by an additional 1.6 kJ.mol(-1) in the presence of the solute. The results seem to indicate that the stabilizing effect is due mainly to a reduction in mobility of the slower, larger-scale motions within the protein structure with an associated increase in the enthalpy of interactions.

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More information

Published date: 1 December 2003
Keywords: chemical exchange, compatible solutes, protein dynamics, rubredoxin, thermostability, backbone dynamics, hydrogen-exchange, preferential interactions, temperature-dependence, globular-proteins, organic solutes, relaxation, stability, binding, spectroscopy

Identifiers

Local EPrints ID: 20006
URI: http://eprints.soton.ac.uk/id/eprint/20006
ISSN: 0014-2956
PURE UUID: 798282d6-e747-46f0-912a-0e8fa4f42892

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Date deposited: 24 Feb 2006
Last modified: 08 Jan 2022 01:00

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Contributors

Author: Pedro Lamosa
Author: David L. Turner
Author: Rita Ventura
Author: Christopher Maycock
Author: Helena Santos

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