Effects of hydrostatic pressure on the quaternary structure and enzymatic activity of a large peptidase complex from Pyrococcus horikoshii

► The TET particle is stable over a large pressure and temperature range. ► TET activity is enhanced by pressure in a non-linear fashion. ► Analysis of the enzymatic parameters suggests a change in the rate-limiting step. While molecular adaptation to high temperature has been extensively studied, t...

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Bibliographic Details
Published in:Archives of biochemistry and biophysics 2012-01, Vol.517 (2), p.104-110
Main Authors: Rosenbaum, Eva, Gabel, Frank, Durá, M. Asunción, Finet, Stéphanie, Cléry-Barraud, Cécile, Masson, Patrick, Franzetti, Bruno
Format: Article
Language:English
Subjects:
Aminopeptidase
Archaeal Proteins - chemistry
Archaeal Proteins - metabolism
catalytic activity
Enzymatic activity
enzyme activity
Enzyme Stability
Hydrostatic Pressure
Hyperthermophiles
Kinetics
Large molecular complexes
Peptide Hydrolases - chemistry
Peptide Hydrolases - metabolism
protein structure
Protein Structure, Quaternary
Pyrococcus horikoshii
Pyrococcus horikoshii - enzymology
Quaternary structure
Recombinant Proteins - chemistry
Recombinant Proteins - metabolism
Scattering, Small Angle
Temperature
X-radiation
X-Ray Diffraction
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Summary:► The TET particle is stable over a large pressure and temperature range. ► TET activity is enhanced by pressure in a non-linear fashion. ► Analysis of the enzymatic parameters suggests a change in the rate-limiting step. While molecular adaptation to high temperature has been extensively studied, the effect of hydrostatic pressure on protein structure and enzymatic activity is still poorly understood. We have studied the influence of pressure on both the quaternary structure and enzymatic activity of the dodecameric TET3 peptidase from Pyrococcus horikoshii. Small angle X-ray scattering (SAXS) revealed a high robustness of the oligomer under high pressure of up to 300 MPa at 25 °C as well as at 90 °C. The enzymatic activity of TET3 was enhanced by pressure up to 180 MPa. From the pressure behavior of the different rate-constants we have determined the volume changes associated with substrate binding and catalysis. Based on these results we propose that a change in the rate-limiting step occurs around 180 MPa.
ISSN:0003-9861
1096-0384
DOI:10.1016/j.abb.2011.07.017