Low barrier hydrogen bond is absent in the catalytic triads in the ground state but is present in a transition-state complex in the prolyl oligopeptidase family of serine proteases

Ara Kahyaoglu, Khadijeh Haghjoo, Fusheng Guo, Frank Jordan, Charles Kettner, Ferenc Felföldi, L. Polgár

Research output: Contribution to journalArticle

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Abstract

High frequency proton NMR spectra for two members of the prolyl oligopeptidase class of serine proteases, prolyl oligopeptidase and oligopeptidase B, showed that resonances corresponding to the active center histidine NS(δ1)H and N(ε2)H generally observed in this region, are absent in these enzymes. However, for both enzymes, as well as with the H652A and H652Q active center variants of oligopeptidase B, there are two resonances observed in this region that could be assigned to two protonated histidines with a noncatalytic function. The results indicate that these two histidines participate in strong hydrogen bonds. The absence of resonances pertinent to the active center histidine resonances suggests the absence of a low barrier hydrogen bond between the Asp and His in these two enzymes in their ground states. Addition of the peptide boronic acid t-butoxycarbonyl-(D)Val-Leu- (L)boroArg to oligopeptidase B resulted in potent, slow binding inhibition of the enzyme and the appearance of a new resonance at 15.8 ppm, whose chemical shift is appropriate for a tetrahedral boronate complex and a low barrier hydrogen bond. The results demonstrate important dissimilarities between the active centers of the prolyl oligopeptidase class of serine proteases and the pancreatic and subtilisin classes both in the ground state and in the transition-state analog complexes.

Original languageEnglish
Pages (from-to)25547-25554
Number of pages8
JournalJournal of Biological Chemistry
Volume272
Issue number41
DOIs
Publication statusPublished - Oct 10 1997

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prolyl oligopeptidase
oligopeptidase B
Serine Proteases
Electron transitions
Histidine
Ground state
Hydrogen
Hydrogen bonds
Enzymes
valylleucine
Boronic Acids
Subtilisin
Protons
Chemical shift
Peptides
Nuclear magnetic resonance

ASJC Scopus subject areas

  • Biochemistry

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Low barrier hydrogen bond is absent in the catalytic triads in the ground state but is present in a transition-state complex in the prolyl oligopeptidase family of serine proteases. / Kahyaoglu, Ara; Haghjoo, Khadijeh; Guo, Fusheng; Jordan, Frank; Kettner, Charles; Felföldi, Ferenc; Polgár, L.

In: Journal of Biological Chemistry, Vol. 272, No. 41, 10.10.1997, p. 25547-25554.

Research output: Contribution to journalArticle

Kahyaoglu, Ara ; Haghjoo, Khadijeh ; Guo, Fusheng ; Jordan, Frank ; Kettner, Charles ; Felföldi, Ferenc ; Polgár, L. / Low barrier hydrogen bond is absent in the catalytic triads in the ground state but is present in a transition-state complex in the prolyl oligopeptidase family of serine proteases. In: Journal of Biological Chemistry. 1997 ; Vol. 272, No. 41. pp. 25547-25554.
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