Ligand-induced conformational changes in prolyl oligopeptidase: a kinetic approach

Abstract Most kinetic studies of prolyl oligopeptidase (PREP) were performed with the porcine enzyme using modified peptide substrates. Yet recent biophysical studies used the human homolog. Therefore, the aim of this study was to compare the kinetic behavior of human and porcine PREP, as well as to...

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Bibliographic Details
Published in:Protein engineering, design and selection design and selection, 2017-03, Vol.30 (3), p.219-226
Main Authors: Van Elzen, R., Schoenmakers, E., Brandt, I., Van Der Veken, P., Lambeir, A.M.
Format: Article
Language:English
Subjects:
Animals
Enzyme Stability
Humans
Hydrogen-Ion Concentration
Kinetics
Mitochondrial Proteins - chemistry
Mitochondrial Proteins - metabolism
Protein Domains
Recombinant Proteins - chemistry
Recombinant Proteins - metabolism
Serine Endopeptidases - chemistry
Serine Endopeptidases - metabolism
Swine
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Summary:Abstract Most kinetic studies of prolyl oligopeptidase (PREP) were performed with the porcine enzyme using modified peptide substrates. Yet recent biophysical studies used the human homolog. Therefore, the aim of this study was to compare the kinetic behavior of human and porcine PREP, as well as to find a suitable method to study enzyme kinetics with an unmodified biological substrate. It was found that human PREP behaves identically to the porcine homolog, displaying a double bell-shaped pH profile and a pH-dependent solvent kinetic isotope effect of the kcat/Km, features that set it apart from the related exopeptidase dipeptidyl peptidase IV (DPP IV). However, the empirical temperature coefficient Q10, describing the temperature dependency of the kinetic parameters and the non-linear Arrhenius plot of kcat/Km are common characteristics between PREP and DPP IV. The results also demonstrate the feasibility of microcalorimetry for measuring turn-over of proline containing peptides.
ISSN:1741-0126
1741-0134
DOI:10.1093/protein/gzw079