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Title: Structural and kinetic insights into the mechanism of 5-hydroxyisourate hydrolase from Klebsiella pneumoniae

Abstract

The stereospecific oxidative degradation of uric acid to (S)-allantoin has recently been demonstrated to proceed via two unstable intermediates and requires three separate enzymatic reactions. The second step of this reaction, the conversion of 5-hydroxyisourate (HIU) to 2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline, is catalyzed by HIU hydrolase (HIUH). The high-resolution crystal structure of HIUH from the opportunistic pathogen Klebsiella pneumoniae (KpHIUH) has been determined. KpHIUH is a homotetrameric protein that, based on sequence and structural similarity, belongs to the transthyretin-related protein family. In addition, the steady-state kinetic parameters for this enzyme and four active-site mutants have been measured. These data provide valuable insight into the functional roles of the active-site residues. Based upon the structural and kinetic data, a mechanism is proposed for the KpHIUH-catalyzed reaction.

Authors:
;  [1]
  1. Cornell
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
National Institutes of Health (NIH)
OSTI Identifier:
1019146
Resource Type:
Journal Article
Journal Name:
Acta Crystallogr. D
Additional Journal Information:
Journal Volume: 67; Journal Issue: (8) ; 08, 2011; Journal ID: ISSN 0907-4449
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; 99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE; CRYSTAL STRUCTURE; ENZYMES; FUNCTIONALS; HYDROLASES; KINETICS; KLEBSIELLA; MUTANTS; PATHOGENS; PROTEINS; RESIDUES; URIC ACID

Citation Formats

French, Jarrod B, and Ealick, Steven E. Structural and kinetic insights into the mechanism of 5-hydroxyisourate hydrolase from Klebsiella pneumoniae. United States: N. p., 2011. Web. doi:10.1107/S090744491101746X.
French, Jarrod B, & Ealick, Steven E. Structural and kinetic insights into the mechanism of 5-hydroxyisourate hydrolase from Klebsiella pneumoniae. United States. https://doi.org/10.1107/S090744491101746X
French, Jarrod B, and Ealick, Steven E. 2011. "Structural and kinetic insights into the mechanism of 5-hydroxyisourate hydrolase from Klebsiella pneumoniae". United States. https://doi.org/10.1107/S090744491101746X.
@article{osti_1019146,
title = {Structural and kinetic insights into the mechanism of 5-hydroxyisourate hydrolase from Klebsiella pneumoniae},
author = {French, Jarrod B and Ealick, Steven E},
abstractNote = {The stereospecific oxidative degradation of uric acid to (S)-allantoin has recently been demonstrated to proceed via two unstable intermediates and requires three separate enzymatic reactions. The second step of this reaction, the conversion of 5-hydroxyisourate (HIU) to 2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline, is catalyzed by HIU hydrolase (HIUH). The high-resolution crystal structure of HIUH from the opportunistic pathogen Klebsiella pneumoniae (KpHIUH) has been determined. KpHIUH is a homotetrameric protein that, based on sequence and structural similarity, belongs to the transthyretin-related protein family. In addition, the steady-state kinetic parameters for this enzyme and four active-site mutants have been measured. These data provide valuable insight into the functional roles of the active-site residues. Based upon the structural and kinetic data, a mechanism is proposed for the KpHIUH-catalyzed reaction.},
doi = {10.1107/S090744491101746X},
url = {https://www.osti.gov/biblio/1019146}, journal = {Acta Crystallogr. D},
issn = {0907-4449},
number = (8) ; 08, 2011,
volume = 67,
place = {United States},
year = {Tue Jul 19 00:00:00 EDT 2011},
month = {Tue Jul 19 00:00:00 EDT 2011}
}