Functional and structural characterization of Streptococcus pneumoniae pyruvate kinase involved in fosfomycin resistance

Taguchi, Atsushi; Nakashima, Ryosuke; Nishino, Kunihiko

Journal of Biological Chemistry, 2023, 299(7), 104892

Number of Access:487（2025-07-30 13:31 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/92477

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Title
Functional and structural characterization of Streptococcus pneumoniae pyruvate kinase involved in fosfomycin resistance
Creator
NRID
1000020908686 1000020908686
ORCID
0000-0003-4962-0393 0000-0003-4962-0393
researchmap permalink
atsushi_taguchi atsushi_taguchi
Scopus Author ID
57199647175 57199647175
CreatorName
Taguchi, Atsushi
NRID
1000020379100 1000020379100
researchmap permalink
nakashi nakashi
Scopus Author ID
7005644436 7005644436
CreatorName
Nakashima, Ryosuke
NRID
1000030432438 1000030432438
ORCID
0000-0003-3349-1769 0000-0003-3349-1769
researchmap permalink
read0210124 read0210124
Scopus Author ID
7102938399 7102938399
CreatorName
Nishino, Kunihiko
Subject
allosteric regulation
antibiotic resistance
enzyme kinetics
enzyme structure
fosfomycin
pyruvate kinase
Streptococcus
Description
Taguchi A., Nakashima R., Nishino K.. Functional and structural characterization of Streptococcus pneumoniae pyruvate kinase involved in fosfomycin resistance. Journal of Biological Chemistry 299, 104892 (2023); https://doi.org/10.1016/j.jbc.2023.104892.
Abstract
Glycolysis is the primary metabolic pathway in the strictly fermentative Streptococcus pneumoniae, which is a major human pathogen associated with antibiotic resistance. Pyruvate kinase (PYK) is the last enzyme in this pathway that catalyzes the production of pyruvate from phosphoenolpyruvate (PEP) and plays a crucial role in controlling carbon flux; however, while S. pneumoniae PYK (SpPYK) is indispensable for growth, surprisingly little is known about its functional properties. Here, we report that compromising mutations in SpPYK confers resistance to the antibiotic fosfomycin, which inhibits the peptidoglycan synthesis enzyme MurA, implying a direct link between PYK and cell wall biogenesis. The crystal structures of SpPYK in the apo and ligand-bound states reveal key interactions that contribute to its conformational change as well as residues responsible for the recognition of PEP and the allosteric activator fructose 1,6-bisphosphate (FBP). Strikingly, FBP binding was observed at a location distinct from previously reported PYK effector binding sites. Furthermore, we show that SpPYK could be engineered to become more responsive to glucose 6-phosphate instead of FBP by sequence and structure-guided mutagenesis of the effector binding site. Together, our work sheds light on the regulatory mechanism of SpPYK and lays the groundwork for antibiotic development that targets this essential enzyme.
Publisher
American Society for Biochemistry and Molecular Biology Inc.
Source Title
Journal of Biological Chemistry
Volume (Issue)
299(7)
Page
104892
Date of Issued
2023-07-01
Language
English
Handle URL
https://hdl.handle.net/11094/92477
PISSN
00219258
eISSN
1083351X
NCID
AA1202441X
Relation.isIdenticalTo
DOI (Publisher Version)
https://doi.org/10.1016/j.jbc.2023.104892
Access Rights
open access
CopyRight
This article is licensed under a Creative Commons Attribution 4.0 International License.
oaire:version
Version of Record
Category
学術雑誌論文 Journal Article