Structural Analysis and Molecular Dynamics Simulations of Urease From Ureaplasma parvum.

Λεπτομέρειες βιβλιογραφικής εγγραφής
Τίτλος: Structural Analysis and Molecular Dynamics Simulations of Urease From Ureaplasma parvum.
Συγγραφείς: Wu HN; Department of Developmental Medicine, Research Institute, Osaka Women's and Children's Hospital, Izumi City 594-1101 Osaka, Japan., Fujita J; Graduate School of Frontier Biosciences, The University of Osaka, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan; JEOL YOKOGUSHI Research Alliance Laboratories, The University of Osaka, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan; Graduate School of Pharmaceutical Sciences, The University of Osaka, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan., Nakura Y; Department of Developmental Medicine, Research Institute, Osaka Women's and Children's Hospital, Izumi City 594-1101 Osaka, Japan., Inoue M; Graduate School of Medical Life Science, Yokohama City University, 1-7-29, Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan., Suzuki K; The Research Foundation for Microbial Diseases of Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan; BIKEN-RIMD NGS Laboratory, Research Institute for Microbial Diseases, The University of Osaka, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan., Ekimoto T; Graduate School of Medical Life Science, Yokohama City University, 1-7-29, Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan., Yin B; Department of Developmental Medicine, Research Institute, Osaka Women's and Children's Hospital, Izumi City 594-1101 Osaka, Japan; Department of Pediatric and Neonatal-Perinatal Research, Affiliate Graduate School of Medicine, The University of Osaka, 2-2 Yamadaoka, Suita 565-0871 Osaka, Japan., Fukuda Y; Graduate School of Pharmaceutical Sciences, The University of Osaka, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan; Integrated Frontier Research for Medical Science Division, Institute for Open and Transdisciplinary Research Initiatives (OTRI), The University of Osaka, Japan., Harada K; Graduate School of Pharmaceutical Sciences, The University of Osaka, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan., Inoue T; Graduate School of Pharmaceutical Sciences, The University of Osaka, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan; Integrated Frontier Research for Medical Science Division, Institute for Open and Transdisciplinary Research Initiatives (OTRI), The University of Osaka, Japan., Ikeguchi M; Graduate School of Medical Life Science, Yokohama City University, 1-7-29, Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan; RIKEN Center for Computational Science, 1-7-22, Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan., Namba K; Graduate School of Frontier Biosciences, The University of Osaka, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan; JEOL YOKOGUSHI Research Alliance Laboratories, The University of Osaka, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan., Yanagihara I; Department of Developmental Medicine, Research Institute, Osaka Women's and Children's Hospital, Izumi City 594-1101 Osaka, Japan; Department of Pediatric and Neonatal-Perinatal Research, Affiliate Graduate School of Medicine, The University of Osaka, 2-2 Yamadaoka, Suita 565-0871 Osaka, Japan. Electronic address: itaruy@wch.opho.jp.
Πηγή: Journal of molecular biology [J Mol Biol] 2025 Nov 01; Vol. 437 (21), pp. 169368. Date of Electronic Publication: 2025 Aug 05.
Τύπος έκδοσης: Journal Article
Γλώσσα: English
Στοιχεία περιοδικού: Publisher: Elsevier Country of Publication: Netherlands NLM ID: 2985088R Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1089-8638 (Electronic) Linking ISSN: 00222836 NLM ISO Abbreviation: J Mol Biol Subsets: MEDLINE
Imprint Name(s): Publication: Amsterdam : Elsevier
Original Publication: 1959- : London : Academic Press
Ιατρικοί όροι (MeSH): Urease*/chemistry , Urease*/metabolism , Urease*/genetics , Ureaplasma*/enzymology, Bacterial Proteins/chemistry ; Bacterial Proteins/metabolism ; Molecular Dynamics Simulation ; Cryoelectron Microscopy ; Catalytic Domain ; Kinetics ; Protein Conformation
Περίληψη: Ureaplasma is one of the smallest pathogenic bacteria, generating approximately 95% of its adenosine triphosphate (ATP) solely through urease. Studies on Ureaplasma parvum, a species of Ureaplasma, have confirmed that adding urease inhibitors inhibits bacterial growth. The Km and Vmax of the urease-mediated reaction were estimated to be 4.3 ± 0.2 mM and 3,333.3 ± 38.0 μmol NH3/min/mg protein, respectively. The cryo-electron microscopy (cryo-EM) structure of Ureaplasma parvum urease (UPU) at a resolution of 2.03 Å reveals a trimer of heterotrimers comprising three proteins: UreA, UreB, and UreC. The active site is well conserved among the known ureases. However, the Vmax of UPU was higher than that of most known ureases, including those ureases derived from Sporosarcina pasteurii (SPU) and Klebsiella aerogenes (KAU) with identical oligomeric state. All-atom molecular dynamics simulations showed that the flap and UreB are more open in UPU than SPU and KAU. His-tagged wild-type recombinant UPU (WT-rUPU) revealed estimated Km and Vmax values of 4.1 ± 0.3 mM and 769.2 ± 7.4 µmol NH3/min/mg protein, respectively. Amino acid substitutions of recombinant UPUs within the flap region to SPU. Amongst the flap region variants, the Vmax of K331N variant was 48-fold lower than that of WT-rUPU. ICP-MS analysis reveals that one molecule of UPU, WT-rUPU, and K331N-rUPU contains 3.7, 0.8, and 0.1 Ni2+ atoms, respectively, suggesting that a wide-open flap of urease may contribute to delivering nickel into the enzyme, resulting in a high Vmax. Ureaplasma evolved highly efficient UPU through a few amino acid substitutions in the disorganized loop of the mobile flap region.
(Copyright © 2025 The Author(s). Published by Elsevier Ltd.. All rights reserved.)
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Contributed Indexing: Keywords: Ureaplasma; cryo-EM; mobile flap; molecular dynamics; urease
Substance Nomenclature: EC 3.5.1.5 (Urease)
0 (Bacterial Proteins)
Entry Date(s): Date Created: 20250802 Date Completed: 20251001 Latest Revision: 20251001
Update Code: 20260130
DOI: 10.1016/j.jmb.2025.169368
PMID: 40752870
Βάση Δεδομένων: MEDLINE
Περιγραφή
ISSN:1089-8638
DOI:10.1016/j.jmb.2025.169368