Truncated prolyl oligopeptidase from Pyrococcus furiosus

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The peptidase domain of prolyl oligopeptidase is covered by a propeller domain, which excludes large peptides and proteins from the catalytic triad. Previous studies indicated that some amino acids of the N-terminal region constitute a part of the substrate entrance to the active site. To investigate the catalytic role of the N-terminus, we removed the residues 1-32 from the enzyme and examined the kinetic, thermodynamic, and structural consequences of the deletion, using the thermophile Pyrococcus furiosus prolyl oligopeptidase. An about threefold decrease in the catalytic activity along with a 20°C reduction in the temperature optimum was observed. The pH-rate profile, the rate-limiting step, and the activation parameters did not change significantly. However, a substantial decrease was observed in the stability of the protein as demonstrated by circular dichroism and differential scanning calorimetry measurements, and by denaturation with guanidinium chloride. It was concluded that the N-terminal segment did not facilitate the substrate binding independent of the size of the substrate, but contributed principally to the protein stability required for the formation of the proper active site.

Original languageEnglish
Pages (from-to)633-643
Number of pages11
JournalProteins: Structure, Function and Genetics
Volume69
Issue number3
DOIs
Publication statusPublished - Nov 15 2007

Fingerprint

prolyl oligopeptidase
Pyrococcus furiosus
Protein Stability
Catalytic Domain
Guanidine
Differential Scanning Calorimetry
Substrates
Circular Dichroism
Thermodynamics
Proteins
Denaturation
Peptide Hydrolases
Propellers
Amino Acids
Peptides
Temperature
Differential scanning calorimetry
Catalyst activity
Enzymes
Chemical activation

Keywords

  • Kinetic analysis
  • Protein stability
  • Refolding
  • Site specific mutagenesis
  • Thermophilic enzyme

ASJC Scopus subject areas

  • Genetics
  • Structural Biology
  • Biochemistry

Cite this

Truncated prolyl oligopeptidase from Pyrococcus furiosus. / Juhász, T.; Szeltner, Z.; Polgár, L.

In: Proteins: Structure, Function and Genetics, Vol. 69, No. 3, 15.11.2007, p. 633-643.

Research output: Contribution to journalArticle

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